BioMEMS Lab

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Journal Publications

152 entries « ‹ 3 of 8 › »

41.	Zhang, Y; Chen, L; Hsieh, K; Wang, T H: Ratiometric Fluorescence Coding for Multiplex Nucleic Acid Amplification Testing. In: Anal Chem, 90 (20), pp. 12180-12186, 2018, ISSN: 1520-6882 (Electronic) 0003-2700 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN6736, title = {Ratiometric Fluorescence Coding for Multiplex Nucleic Acid Amplification Testing}, author = { Y. Zhang and L. Chen and K. Hsieh and T. H. Wang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/30251853}, doi = {10.1021/acs.analchem.8b03266}, issn = {1520-6882 (Electronic) 0003-2700 (Linking)}, year = {2018}, date = {2018-10-16}, journal = {Anal Chem}, volume = {90}, number = {20}, pages = {12180-12186}, abstract = {Although nucleic acid amplification testing (NAAT) has become the cornerstone for molecular diagnosis of diseases, expanding the multiplexed detection capacity of NAAT remains an important objective. To this end, encoding each nucleic acid target with a specific fluorescently labeled probe has been the most mature approach for multiplexed detection. Unfortunately, the number of targets that can be differentiated via this one-target-one-fluorophore multiplexed detection approach is restricted by spectral overlaps between fluorophores. In response, we present herein a new multiplexed detection approach termed ratiometric fluorescence coding, in which we encode each nucleic acid target with a specific ratio between two standard fluorophores. In ratiometric fluorescence coding, we employ the padlock probe chemistry to encode each nucleic acid target with a specific number of binding sites for two probes labeled with different fluorophores. Coupling the padlock probes with either rolling circle amplification (RCA) or hyperbranched rolling circle amplification (HRCA), we transform each nucleic acid target into a specific template that allows hybridization with the fluorescently labeled probes at predesigned ratios, thereby achieving multiplexed detection. For demonstration, we detected DNA targets from six infectious diseases and demonstrated the potential for further expanding the multiplexing capability of our approach. With further development, ratiometric fluorescence coding has the potential to enable highly multiplexed detection of nucleic acid targets and facilitate molecular diagnosis of diseases.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Although nucleic acid amplification testing (NAAT) has become the cornerstone for molecular diagnosis of diseases, expanding the multiplexed detection capacity of NAAT remains an important objective. To this end, encoding each nucleic acid target with a specific fluorescently labeled probe has been the most mature approach for multiplexed detection. Unfortunately, the number of targets that can be differentiated via this one-target-one-fluorophore multiplexed detection approach is restricted by spectral overlaps between fluorophores. In response, we present herein a new multiplexed detection approach termed ratiometric fluorescence coding, in which we encode each nucleic acid target with a specific ratio between two standard fluorophores. In ratiometric fluorescence coding, we employ the padlock probe chemistry to encode each nucleic acid target with a specific number of binding sites for two probes labeled with different fluorophores. Coupling the padlock probes with either rolling circle amplification (RCA) or hyperbranched rolling circle amplification (HRCA), we transform each nucleic acid target into a specific template that allows hybridization with the fluorescently labeled probes at predesigned ratios, thereby achieving multiplexed detection. For demonstration, we detected DNA targets from six infectious diseases and demonstrated the potential for further expanding the multiplexing capability of our approach. With further development, ratiometric fluorescence coding has the potential to enable highly multiplexed detection of nucleic acid targets and facilitate molecular diagnosis of diseases. Close https://www.ncbi.nlm.nih.gov/pubmed/30251853 doi:10.1021/acs.analchem.8b03266 Close
42.	Andini, N; Hu, A; Zhou, L; Cogill, S; Wang, T H; Wittwer, C T; Yang, S: A "Culture" Shift: Broad Bacterial Detection, Identification, and Antimicrobial Susceptibility Testing Directly from Whole Blood. In: Clin Chem, 64 (10), pp. 1453-1462, 2018, ISSN: 1530-8561 (Electronic) 0009-9147 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN6607, title = {A "Culture" Shift: Broad Bacterial Detection, Identification, and Antimicrobial Susceptibility Testing Directly from Whole Blood}, author = { N. Andini and A. Hu and L. Zhou and S. Cogill and T. H. Wang and C. T. Wittwer and S. Yang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/30087140}, doi = {10.1373/clinchem.2018.290189}, issn = {1530-8561 (Electronic) 0009-9147 (Linking)}, year = {2018}, date = {2018-10-01}, journal = {Clin Chem}, volume = {64}, number = {10}, pages = {1453-1462}, abstract = {BACKGROUND: The time required for bloodstream pathogen detection, identification (ID), and antimicrobial susceptibility testing (AST) does not satisfy the acute needs of disease management. Conventional methods take up to 3 days for ID and AST. Molecular diagnostics have reduced times for ID, but their promise to supplant culture is unmet because AST times remain slow. We developed a combined quantitative PCR (qPCR)-based ID+AST assay with sequential detection, ID, and AST of leading nosocomial bacterial pathogens. METHODS: ID+AST was performed on whole blood samples by (a) removing blood cells, (b) brief bacterial enrichment, (c) bacterial detection and ID, and (d) species-specific antimicrobial treatment. Broad-spectrum qPCR of the internal transcribed spacer between the 16S and 23S was amplified for detection. High-resolution melting identified the species with a curve classifier. AST was enabled by Ct differences between treated and untreated samples. RESULTS: A detection limit of 1 CFU/mL was achieved for Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus. All species were accurately identified by unique melting curves. Antimicrobial minimum inhibitory concentrations were identified with Ct differences of >/=1 cycle. Using an RNA target allowed reduction of AST incubation time from 60 min to 5 min. Rapid-cycle amplification reduced qPCR times by 83% to 30 min. CONCLUSIONS: Combined, sequential ID+AST protocols allow rapid and reliable detection, ID, and AST for the diagnosis of bloodstream infections, enabling conversion of empiric to targeted therapy by the second dose of antimicrobials.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close BACKGROUND: The time required for bloodstream pathogen detection, identification (ID), and antimicrobial susceptibility testing (AST) does not satisfy the acute needs of disease management. Conventional methods take up to 3 days for ID and AST. Molecular diagnostics have reduced times for ID, but their promise to supplant culture is unmet because AST times remain slow. We developed a combined quantitative PCR (qPCR)-based ID+AST assay with sequential detection, ID, and AST of leading nosocomial bacterial pathogens. METHODS: ID+AST was performed on whole blood samples by (a) removing blood cells, (b) brief bacterial enrichment, (c) bacterial detection and ID, and (d) species-specific antimicrobial treatment. Broad-spectrum qPCR of the internal transcribed spacer between the 16S and 23S was amplified for detection. High-resolution melting identified the species with a curve classifier. AST was enabled by Ct differences between treated and untreated samples. RESULTS: A detection limit of 1 CFU/mL was achieved for Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus. All species were accurately identified by unique melting curves. Antimicrobial minimum inhibitory concentrations were identified with Ct differences of >/=1 cycle. Using an RNA target allowed reduction of AST incubation time from 60 min to 5 min. Rapid-cycle amplification reduced qPCR times by 83% to 30 min. CONCLUSIONS: Combined, sequential ID+AST protocols allow rapid and reliable detection, ID, and AST for the diagnosis of bloodstream infections, enabling conversion of empiric to targeted therapy by the second dose of antimicrobials. Close https://www.ncbi.nlm.nih.gov/pubmed/30087140 doi:10.1373/clinchem.2018.290189 Close
43.	Chen, L; Shin, D J; Zheng, S; Melendez, J H; Gaydos, C A; Wang, T H: Direct-qPCR Assay for Coupled Identification and Antimicrobial Susceptibility Testing of Neisseria gonorrhoeae. In: ACS Infect Dis, 4 (9), pp. 1377-1384, 2018, ISSN: 2373-8227 (Electronic) 2373-8227 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN6641, title = {Direct-qPCR Assay for Coupled Identification and Antimicrobial Susceptibility Testing of Neisseria gonorrhoeae}, author = { L. Chen and D. J. Shin and S. Zheng and J. H. Melendez and C. A. Gaydos and T. H. Wang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/29999304}, doi = {10.1021/acsinfecdis.8b00104}, issn = {2373-8227 (Electronic) 2373-8227 (Linking)}, year = {2018}, date = {2018-09-14}, journal = {ACS Infect Dis}, volume = {4}, number = {9}, pages = {1377-1384}, abstract = {Multidrug-resistant gonorrhea has become an urgent issue for global public health. As the causative agent of gonorrhea, Neisseria gonorrhoeae, has been progressively developing resistance to nearly all prescribed antimicrobial drugs, monitoring its antimicrobial resistance on a broader scale has become a crucial agenda for effective antibiotic stewardship. Unfortunately, gold standard antimicrobial susceptibility testing (AST) relies on time and labor-intensive phenotypic assays, which lag behind the current diagnostic workflow for N. gonorrhoeae identification based on nucleic acid amplification tests (NAAT). Newer assay technologies based on NAAT can rapidly identify N. gonorrhoeae from clinical specimen but fundamentally lack the capacity to provide phenotypic AST information. Herein, we propose a direct-quantitative PCR (direct-qPCR) assay that enables pathogen-specific identification and phenotypic AST via quantitative measurement of N. gonorrhoeae growth directly from a liquid medium without any sample preprocessing. The assay has an analytical sensitivity of 10(2) CFU/mL and is highly specific to N. gonorrhoeae in the presence of urogenital flora and clinical swab eluent. We tested seven N. gonorrhoeae strains against three antibiotic agents, penicillin, tetracycline, and ciprofloxacin, and achieved 95.2% category agreement and 85.7% essential agreement with the FDA-approved E-test. The assay presented in this work has the unique ability to identify N. gonorrhoeae and provide phenotypic AST directly from the liquid medium with cell densities as low as 10(2) CFU/mL, demonstrating an accelerated, sensitive, and scalable workflow for performing both identification and AST of N. gonorrhoeae.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Multidrug-resistant gonorrhea has become an urgent issue for global public health. As the causative agent of gonorrhea, Neisseria gonorrhoeae, has been progressively developing resistance to nearly all prescribed antimicrobial drugs, monitoring its antimicrobial resistance on a broader scale has become a crucial agenda for effective antibiotic stewardship. Unfortunately, gold standard antimicrobial susceptibility testing (AST) relies on time and labor-intensive phenotypic assays, which lag behind the current diagnostic workflow for N. gonorrhoeae identification based on nucleic acid amplification tests (NAAT). Newer assay technologies based on NAAT can rapidly identify N. gonorrhoeae from clinical specimen but fundamentally lack the capacity to provide phenotypic AST information. Herein, we propose a direct-quantitative PCR (direct-qPCR) assay that enables pathogen-specific identification and phenotypic AST via quantitative measurement of N. gonorrhoeae growth directly from a liquid medium without any sample preprocessing. The assay has an analytical sensitivity of 10(2) CFU/mL and is highly specific to N. gonorrhoeae in the presence of urogenital flora and clinical swab eluent. We tested seven N. gonorrhoeae strains against three antibiotic agents, penicillin, tetracycline, and ciprofloxacin, and achieved 95.2% category agreement and 85.7% essential agreement with the FDA-approved E-test. The assay presented in this work has the unique ability to identify N. gonorrhoeae and provide phenotypic AST directly from the liquid medium with cell densities as low as 10(2) CFU/mL, demonstrating an accelerated, sensitive, and scalable workflow for performing both identification and AST of N. gonorrhoeae. Close https://www.ncbi.nlm.nih.gov/pubmed/29999304 doi:10.1021/acsinfecdis.8b00104 Close
44.	O'Keefe, C M; Pisanic, 2nd T R; Zec, H; Overman, M J; Herman, J G; Wang, T H: Facile profiling of molecular heterogeneity by microfluidic digital melt. In: Sci Adv, 4 (9), pp. eaat6459, 2018, ISSN: 2375-2548 (Electronic) 2375-2548 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN6702, title = {Facile profiling of molecular heterogeneity by microfluidic digital melt}, author = { C. M. O'Keefe and T. R. 2nd Pisanic and H. Zec and M. J. Overman and J. G. Herman and T. H. Wang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/30263958}, doi = {10.1126/sciadv.aat6459}, issn = {2375-2548 (Electronic) 2375-2548 (Linking)}, year = {2018}, date = {2018-09-01}, journal = {Sci Adv}, volume = {4}, number = {9}, pages = {eaat6459}, abstract = {This work presents a digital microfluidic platform called HYPER-Melt (high-density profiling and enumeration by melt) for highly parallelized copy-by-copy DNA molecular profiling. HYPER-Melt provides a facile means of detecting and assessing sequence variations of thousands of individual DNA molecules through digitization in a nanowell microchip array, allowing amplification and interrogation of individual template molecules by detecting HRM fluorescence changes due to sequence-dependent denaturation. As a model application, HYPER-Melt is used here for the detection and assessment of intermolecular heterogeneity of DNA methylation within the promoters of classical tumor suppressor genes. The capabilities of this platform are validated through serial dilutions of mixed epialleles, with demonstrated detection limits as low as 1 methylated variant in 2 million unmethylated templates (0.00005%) of a classic tumor suppressor gene, CDKN2A (p14(ARF)). The clinical potential of the platform is demonstrated using a digital assay for NDRG4, a tumor suppressor gene that is commonly methylated in colorectal cancer, in liquid biopsies of healthy and colorectal cancer patients. Overall, the platform provides the depth of information, simplicity of use, and single-molecule sensitivity necessary for rapid assessment of intermolecular variation contributing to genetic and epigenetic heterogeneity for challenging applications in embryogenesis, carcinogenesis, and rare biomarker detection.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close This work presents a digital microfluidic platform called HYPER-Melt (high-density profiling and enumeration by melt) for highly parallelized copy-by-copy DNA molecular profiling. HYPER-Melt provides a facile means of detecting and assessing sequence variations of thousands of individual DNA molecules through digitization in a nanowell microchip array, allowing amplification and interrogation of individual template molecules by detecting HRM fluorescence changes due to sequence-dependent denaturation. As a model application, HYPER-Melt is used here for the detection and assessment of intermolecular heterogeneity of DNA methylation within the promoters of classical tumor suppressor genes. The capabilities of this platform are validated through serial dilutions of mixed epialleles, with demonstrated detection limits as low as 1 methylated variant in 2 million unmethylated templates (0.00005%) of a classic tumor suppressor gene, CDKN2A (p14(ARF)). The clinical potential of the platform is demonstrated using a digital assay for NDRG4, a tumor suppressor gene that is commonly methylated in colorectal cancer, in liquid biopsies of healthy and colorectal cancer patients. Overall, the platform provides the depth of information, simplicity of use, and single-molecule sensitivity necessary for rapid assessment of intermolecular variation contributing to genetic and epigenetic heterogeneity for challenging applications in embryogenesis, carcinogenesis, and rare biomarker detection. Close https://www.ncbi.nlm.nih.gov/pubmed/30263958 doi:10.1126/sciadv.aat6459 Close
45.	Hsieh, K; Zec, H C; Chen, L; Kaushik, A M; Mach, K E; Liao, J C; Wang, T H: Simple and Precise Counting of Viable Bacteria by Resazurin-Amplified Picoarray Detection. In: Anal Chem, 90 (15), pp. 9449-9456, 2018, ISSN: 1520-6882 (Electronic) 0003-2700 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN6643, title = {Simple and Precise Counting of Viable Bacteria by Resazurin-Amplified Picoarray Detection}, author = { K. Hsieh and H. C. Zec and L. Chen and A. M. Kaushik and K. E. Mach and J. C. Liao and T. H. Wang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/29969556}, doi = {10.1021/acs.analchem.8b02096}, issn = {1520-6882 (Electronic) 0003-2700 (Linking)}, year = {2018}, date = {2018-08-07}, journal = {Anal Chem}, volume = {90}, number = {15}, pages = {9449-9456}, abstract = {Simple, fast, and precise counting of viable bacteria is fundamental to a variety of microbiological applications such as food quality monitoring and clinical diagnosis. To this end, agar plating, microscopy, and emerging microfluidic devices for single bacteria detection have provided useful means for counting viable bacteria, but they also have their limitations ranging from complexity, time, and inaccuracy. We present herein our new method RAPiD (Resazurin-Amplified Picoarray Detection) for addressing this important problem. In RAPiD, we employ vacuum-assisted sample loading and oil-driven sample digitization to stochastically confine single bacteria in Picoarray, a microfluidic device with picoliter-sized isolation chambers (picochambers), in <30 s with only a few minutes of hands-on time. We add AlamarBlue, a resazurin-based fluorescent dye for bacterial growth, in our assay to accelerate the detection of "microcolonies" proliferated from single bacteria within picochambers. Detecting fluorescence in picochambers as an amplified surrogate for bacterial cells allows us to count hundreds of microcolonies with a single image taken via wide-field fluorescence microscopy. We have also expanded our method to practically test multiple titrations from a single bacterial sample in parallel. Using this expanded "multi-RAPiD" strategy, we can quantify viable cells in E. coli and S. aureus samples with precision in approximately 3 h, illustrating RAPiD as a promising new method for counting viable bacteria for microbiological applications.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Simple, fast, and precise counting of viable bacteria is fundamental to a variety of microbiological applications such as food quality monitoring and clinical diagnosis. To this end, agar plating, microscopy, and emerging microfluidic devices for single bacteria detection have provided useful means for counting viable bacteria, but they also have their limitations ranging from complexity, time, and inaccuracy. We present herein our new method RAPiD (Resazurin-Amplified Picoarray Detection) for addressing this important problem. In RAPiD, we employ vacuum-assisted sample loading and oil-driven sample digitization to stochastically confine single bacteria in Picoarray, a microfluidic device with picoliter-sized isolation chambers (picochambers), in <30 s with only a few minutes of hands-on time. We add AlamarBlue, a resazurin-based fluorescent dye for bacterial growth, in our assay to accelerate the detection of "microcolonies" proliferated from single bacteria within picochambers. Detecting fluorescence in picochambers as an amplified surrogate for bacterial cells allows us to count hundreds of microcolonies with a single image taken via wide-field fluorescence microscopy. We have also expanded our method to practically test multiple titrations from a single bacterial sample in parallel. Using this expanded "multi-RAPiD" strategy, we can quantify viable cells in E. coli and S. aureus samples with precision in approximately 3 h, illustrating RAPiD as a promising new method for counting viable bacteria for microbiological applications. Close https://www.ncbi.nlm.nih.gov/pubmed/29969556 doi:10.1021/acs.analchem.8b02096 Close
46.	Surrette, C; Scherer, B; Corwin, A; Grossmann, G; Kaushik, A M; Hsieh, K; Zhang, P; Liao, J C; Wong, P K; Wang, T H; Puleo, C M: Rapid Microbiology Screening in Pharmaceutical Workflows. In: SLAS Technol, 23 (4), pp. 387-394, 2018, ISSN: 2472-6311 (Electronic) 2472-6303 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN6700, title = {Rapid Microbiology Screening in Pharmaceutical Workflows}, author = { C. Surrette and B. Scherer and A. Corwin and G. Grossmann and A. M. Kaushik and K. Hsieh and P. Zhang and J. C. Liao and P. K. Wong and T. H. Wang and C. M. Puleo}, url = {https://www.ncbi.nlm.nih.gov/pubmed/30027813}, doi = {10.1177/2472630318779758}, issn = {2472-6311 (Electronic) 2472-6303 (Linking)}, year = {2018}, date = {2018-08-01}, journal = {SLAS Technol}, volume = {23}, number = {4}, pages = {387-394}, abstract = {Recently advances in miniaturization and automation have been utilized to rapidly decrease the time to result for microbiology testing in the clinic. These advances have been made due to the limitations of conventional culture-based microbiology methods, including agar plate and microbroth dilution, which have long turnaround times and require physicians to treat patients empirically with antibiotics before test results are available. Currently, there exist similar limitations in pharmaceutical sterility and bioburden testing, where the long turnaround times associated with standard microbiology testing drive costly inefficiencies in workflows. These include the time lag associated with sterility screening within drug production lines and the warehousing cost and time delays within supply chains during product testing. Herein, we demonstrate a proof-of-concept combination of a rapid microfluidic assay and an efficient cell filtration process that enables a path toward integrating rapid tests directly into pharmaceutical microbiological screening workflows. We demonstrate separation and detection of Escherichia coli directly captured and analyzed from a mammalian (i.e., CHO) cell culture with a 3.0 h incubation. The demonstration is performed using a membrane filtration module that is compatible with sampling from bioreactors, enabling in-line sampling and process monitoring.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Recently advances in miniaturization and automation have been utilized to rapidly decrease the time to result for microbiology testing in the clinic. These advances have been made due to the limitations of conventional culture-based microbiology methods, including agar plate and microbroth dilution, which have long turnaround times and require physicians to treat patients empirically with antibiotics before test results are available. Currently, there exist similar limitations in pharmaceutical sterility and bioburden testing, where the long turnaround times associated with standard microbiology testing drive costly inefficiencies in workflows. These include the time lag associated with sterility screening within drug production lines and the warehousing cost and time delays within supply chains during product testing. Herein, we demonstrate a proof-of-concept combination of a rapid microfluidic assay and an efficient cell filtration process that enables a path toward integrating rapid tests directly into pharmaceutical microbiological screening workflows. We demonstrate separation and detection of Escherichia coli directly captured and analyzed from a mammalian (i.e., CHO) cell culture with a 3.0 h incubation. The demonstration is performed using a membrane filtration module that is compatible with sampling from bioreactors, enabling in-line sampling and process monitoring. Close https://www.ncbi.nlm.nih.gov/pubmed/30027813 doi:10.1177/2472630318779758 Close
47.	Song, Jian; Dailey, Jennifer; Li, Hui; Jang, Hyun-June; Russell, Luisa; Zhang, Pengfei; Searson, Peter C; Wang, Jeff Tza-Huei; Everett, Allen D; Katz, Howard E: Influence of Bioreceptor Layer Structure on Myelin Basic Protein Detection using Organic Field Effect Transistor-Based Biosensors. In: Advanced Functional Materials, 28 (37), pp. 1802605, 2018, ISSN: 1616301X. (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN6701, title = {Influence of Bioreceptor Layer Structure on Myelin Basic Protein Detection using Organic Field Effect Transistor-Based Biosensors}, author = { Jian Song and Jennifer Dailey and Hui Li and Hyun-June Jang and Luisa Russell and Pengfei Zhang and Peter C. Searson and Jeff Tza-Huei Wang and Allen D. Everett and Howard E. Katz}, url = {https://onlinelibrary.wiley.com/doi/abs/10.1002/adfm.201802605}, doi = {10.1002/adfm.201802605}, issn = {1616301X}, year = {2018}, date = {2018-07-31}, journal = {Advanced Functional Materials}, volume = {28}, number = {37}, pages = {1802605}, abstract = {Abstract Organic field-effect transistor (OFET)-based biosensors with antibody receptors are widely considered for protein antigen detection. To the authors' knowledge, there are no comparative evaluations to date of different choices for the matrix polymer to which the antibodies are attached. Herein, multiple acrylic copolymers are studied as receptor layers with myelin basic protein and its corresponding antibody as an archetypal antibody–antigen pair. Stability against multiple washing steps and the capacity for immobilizing antibodies on polymers on device surfaces with the help of fluorescein isothiocyanate-labeled antibodies are compared. Electronic detection and selectivity are also observed. The conclusions provide guidance on the selection of bioreceptor material for increasing sensitivity and process stability of OFET biosensors.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Abstract Organic field-effect transistor (OFET)-based biosensors with antibody receptors are widely considered for protein antigen detection. To the authors' knowledge, there are no comparative evaluations to date of different choices for the matrix polymer to which the antibodies are attached. Herein, multiple acrylic copolymers are studied as receptor layers with myelin basic protein and its corresponding antibody as an archetypal antibody–antigen pair. Stability against multiple washing steps and the capacity for immobilizing antibodies on polymers on device surfaces with the help of fluorescein isothiocyanate-labeled antibodies are compared. Electronic detection and selectivity are also observed. The conclusions provide guidance on the selection of bioreceptor material for increasing sensitivity and process stability of OFET biosensors. Close https://onlinelibrary.wiley.com/doi/abs/10.1002/adfm.201802605 doi:10.1002/adfm.201802605 Close
48.	Shin, D J; Trick, A Y; Hsieh, Y H; Thomas, D L; Wang, T H: Sample-to-Answer Droplet Magnetofluidic Platform for Point-of-Care Hepatitis C Viral Load Quantitation. In: Sci Rep, 8 (1), pp. 9793, 2018, ISSN: 2045-2322 (Electronic) 2045-2322 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN6503, title = {Sample-to-Answer Droplet Magnetofluidic Platform for Point-of-Care Hepatitis C Viral Load Quantitation}, author = { D. J. Shin and A. Y. Trick and Y. H. Hsieh and D. L. Thomas and T. H. Wang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/29955160}, doi = {10.1038/s41598-018-28124-3}, issn = {2045-2322 (Electronic) 2045-2322 (Linking)}, year = {2018}, date = {2018-06-28}, journal = {Sci Rep}, volume = {8}, number = {1}, pages = {9793}, abstract = {Gold standard quantitative nucleic acid tests for diagnosis of viral diseases are currently limited to implementation in laboratories outside of the clinic. An instrument for conducting nucleic acid testing at the point-of-care (POC) that is easily operable by the clinician would reduce the required number of visits to the clinic and improve patient retention for proper treatment. Here we present a droplet magnetofluidic (DM) platform, which leverages functionalized magnetic particles to miniaturize and automate laboratory assays for use in the clinic at the POC. Our novel thermoformed disposable cartridge coupled to a portable multiaxial magnetofluidic instrument enables real-time PCR assays for quantitative and sensitive detection of nucleic acids from crude biosamples. Instead of laborious benchtop sample purification techniques followed by elution and spiking into PCR buffer, the user simply injects the biosample of interest into a cartridge with magnetic particles and loads the cartridge into the instrument. We demonstrate the utility of our platform with hepatitis C virus (HCV) RNA viral load quantitation from blood serum in approximately 1 hour. Clinical serum samples (n = 18) were directly processed on cartridges with no false positives and a limit of detection of 45 IU per 10 microl sample injection.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Gold standard quantitative nucleic acid tests for diagnosis of viral diseases are currently limited to implementation in laboratories outside of the clinic. An instrument for conducting nucleic acid testing at the point-of-care (POC) that is easily operable by the clinician would reduce the required number of visits to the clinic and improve patient retention for proper treatment. Here we present a droplet magnetofluidic (DM) platform, which leverages functionalized magnetic particles to miniaturize and automate laboratory assays for use in the clinic at the POC. Our novel thermoformed disposable cartridge coupled to a portable multiaxial magnetofluidic instrument enables real-time PCR assays for quantitative and sensitive detection of nucleic acids from crude biosamples. Instead of laborious benchtop sample purification techniques followed by elution and spiking into PCR buffer, the user simply injects the biosample of interest into a cartridge with magnetic particles and loads the cartridge into the instrument. We demonstrate the utility of our platform with hepatitis C virus (HCV) RNA viral load quantitation from blood serum in approximately 1 hour. Clinical serum samples (n = 18) were directly processed on cartridges with no false positives and a limit of detection of 45 IU per 10 microl sample injection. Close https://www.ncbi.nlm.nih.gov/pubmed/29955160 doi:10.1038/s41598-018-28124-3 Close
49.	Shin, D J; Lewis, M; Hsieh, Y H; Rahmoun, N A; Gaydos, C A; Wang, T H; Rothman, R: Healthcare worker feedback on a prototype smartphone-based point-of-care test platform for use in episodic care. In: Point Care, 17 (2), pp. 63-65, 2018, ISSN: 1533-029X (Print) 1533-029X (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN6505, title = {Healthcare worker feedback on a prototype smartphone-based point-of-care test platform for use in episodic care}, author = { D. J. Shin and M. Lewis and Y. H. Hsieh and N. A. Rahmoun and C. A. Gaydos and T. H. Wang and R. Rothman}, url = {https://www.ncbi.nlm.nih.gov/pubmed/30245594}, doi = {10.1097/POC.0000000000000160}, issn = {1533-029X (Print) 1533-029X (Linking)}, year = {2018}, date = {2018-06-01}, journal = {Point Care}, volume = {17}, number = {2}, pages = {63-65}, abstract = {With a growing list of new platforms, end-user acceptability is an evolving topic in point-of-care (POC) test development. While technical reports of new experimental POC tests are common, it is rare to find reports which evaluate the end-user acceptability of such innovations. This work illustrates an example of bridging that gap by evaluating the end-user acceptability of an experimental POC test platform with novel technical features. A prototype smartphone-based STI tests was evaluated by ED technicians, followed by a survey of acceptability factors. Our findings suggest that the end-user acceptability of some design features implemented in the prototype.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close With a growing list of new platforms, end-user acceptability is an evolving topic in point-of-care (POC) test development. While technical reports of new experimental POC tests are common, it is rare to find reports which evaluate the end-user acceptability of such innovations. This work illustrates an example of bridging that gap by evaluating the end-user acceptability of an experimental POC test platform with novel technical features. A prototype smartphone-based STI tests was evaluated by ED technicians, followed by a survey of acceptability factors. Our findings suggest that the end-user acceptability of some design features implemented in the prototype. Close https://www.ncbi.nlm.nih.gov/pubmed/30245594 doi:10.1097/POC.0000000000000160 Close
50.	Stark, A; Shin, D J; Wang, T H: A sample-to-answer droplet magnetofluidic assay platform for quantitative methylation-specific PCR. In: Biomed Microdevices, 20 (2), pp. 31, 2018, ISSN: 1572-8781 (Electronic) 1387-2176 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN5843, title = {A sample-to-answer droplet magnetofluidic assay platform for quantitative methylation-specific PCR}, author = { A. Stark and D. J. Shin and T. H. Wang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/29594810}, doi = {10.1007/s10544-018-0276-6}, issn = {1572-8781 (Electronic) 1387-2176 (Linking)}, year = {2018}, date = {2018-03-28}, journal = {Biomed Microdevices}, volume = {20}, number = {2}, pages = {31}, abstract = {Dysregulation of DNA methylation has been identified as an epigenetic biomarker for numerous cancer types. Gene-specific identification techniques relying on methylation-specific PCR (MSP) require a lengthy manual benchtop process that is susceptible to human-error and contamination. This MSP assay requires a series of discrete sample processing steps including genomic DNA extraction, bisulfite conversion and readout via PCR. In this work, we present a streamlined assay platform utilizing droplet magnetofluidic principles for integration of all sample processing steps required to obtain quantitative MSP signal from raw biological samples. We present a streamlined protocol for solid-phase extraction and bisulfite conversion of genomic DNA, which minimizes reagent use and simplifies the sample preparation protocol for implementation on a compact assay platform. Furthermore, we present a thermally robust assay chip that enables DNA extraction, bisulfite conversion and quantitative PCR from biological samples on a single device. Technical improvements to facilitate DNA extraction and PCR on a single chip in addition to chip performance characterization data are presented.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Dysregulation of DNA methylation has been identified as an epigenetic biomarker for numerous cancer types. Gene-specific identification techniques relying on methylation-specific PCR (MSP) require a lengthy manual benchtop process that is susceptible to human-error and contamination. This MSP assay requires a series of discrete sample processing steps including genomic DNA extraction, bisulfite conversion and readout via PCR. In this work, we present a streamlined assay platform utilizing droplet magnetofluidic principles for integration of all sample processing steps required to obtain quantitative MSP signal from raw biological samples. We present a streamlined protocol for solid-phase extraction and bisulfite conversion of genomic DNA, which minimizes reagent use and simplifies the sample preparation protocol for implementation on a compact assay platform. Furthermore, we present a thermally robust assay chip that enables DNA extraction, bisulfite conversion and quantitative PCR from biological samples on a single device. Technical improvements to facilitate DNA extraction and PCR on a single chip in addition to chip performance characterization data are presented. Close https://www.ncbi.nlm.nih.gov/pubmed/29594810 doi:10.1007/s10544-018-0276-6 Close
51.	Zec, Helena C; Zheng, Tony; Liu, Lingshu; Hsieh, Kuangwen; Rane, Tushar D; Pederson, Todd; Wang, Tza-Huei: Programmable microfluidic genotyping of plant DNA samples for marker-assisted selection. In: Microsystems & Nanoengineering, 4 , pp. 17097, 2018, ISSN: 2055-7434. (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN5842, title = {Programmable microfluidic genotyping of plant DNA samples for marker-assisted selection}, author = { Helena C. Zec and Tony Zheng and Lingshu Liu and Kuangwen Hsieh and Tushar D. Rane and Todd Pederson and Tza-Huei Wang}, url = {http://dx.doi.org/10.1038/micronano.2017.97}, doi = {10.1038/micronano.2017.97}, issn = {2055-7434}, year = {2018}, date = {2018-03-26}, journal = {Microsystems & Nanoengineering}, volume = {4}, pages = {17097}, abstract = {As demands to maintain the global food production continue to mount, multinational seed companies are turning to new DNA marker technologies to accelerate the rate of plant breeding and crop improvement. The key to widespread adoption of molecular breeding is the availability of flexible and cost-effective tools that can perform combinatorial and high-throughput genotyping of single-nucleotide polymorphisms (SNPs) to guide the crop development process. Toward this end, we have developed a programmable, droplet-based microfluidic device for genotyping maize genomic DNA. A unique feature of the microfluidic platform is the nano sample processors (NSPs), which allow the device to sequentially load an unrestricted number of unique DNA samples using only two inlets, overcoming the current limitation to the number of sample inputs due to small device footprint. Direct and programmable droplet generation within the device allows each sample to be genotyped against a panel of markers on demand. Moreover, we have successfully implemented the Invader assay for SNP genotyping in flowing, 50-nL droplets, thus achieving significant reduction in consumption of reagents per reaction as compared with conventional genotyping platforms. As a demonstration, we performed 240 Invader reactions (testing 8 DNA samples against 10 SNP markers) and achieved greater than 93% accuracy in SNP calling of plant DNA samples in a single droplet-based experiment.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close As demands to maintain the global food production continue to mount, multinational seed companies are turning to new DNA marker technologies to accelerate the rate of plant breeding and crop improvement. The key to widespread adoption of molecular breeding is the availability of flexible and cost-effective tools that can perform combinatorial and high-throughput genotyping of single-nucleotide polymorphisms (SNPs) to guide the crop development process. Toward this end, we have developed a programmable, droplet-based microfluidic device for genotyping maize genomic DNA. A unique feature of the microfluidic platform is the nano sample processors (NSPs), which allow the device to sequentially load an unrestricted number of unique DNA samples using only two inlets, overcoming the current limitation to the number of sample inputs due to small device footprint. Direct and programmable droplet generation within the device allows each sample to be genotyped against a panel of markers on demand. Moreover, we have successfully implemented the Invader assay for SNP genotyping in flowing, 50-nL droplets, thus achieving significant reduction in consumption of reagents per reaction as compared with conventional genotyping platforms. As a demonstration, we performed 240 Invader reactions (testing 8 DNA samples against 10 SNP markers) and achieved greater than 93% accuracy in SNP calling of plant DNA samples in a single droplet-based experiment. Close http://dx.doi.org/10.1038/micronano.2017.97 doi:10.1038/micronano.2017.97 Close
52.	Kaushik, A M; Hsieh, K; Chen, L; Shin, D J; Liao, J C; Wang, T H: Accelerating bacterial growth detection and antimicrobial susceptibility assessment in integrated picoliter droplet platform. In: Biosens Bioelectron, 97 , pp. 260-266, 2017, ISSN: 1873-4235 (Electronic) 0956-5663 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN5631, title = {Accelerating bacterial growth detection and antimicrobial susceptibility assessment in integrated picoliter droplet platform}, author = { A. M. Kaushik and K. Hsieh and L. Chen and D. J. Shin and J. C. Liao and T. H. Wang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/28609716}, doi = {10.1016/j.bios.2017.06.006}, issn = {1873-4235 (Electronic) 0956-5663 (Linking)}, year = {2017}, date = {2017-11-15}, journal = {Biosens Bioelectron}, volume = {97}, pages = {260-266}, abstract = {There remains an urgent need for rapid diagnostic methods that can evaluate antibiotic resistance for pathogenic bacteria in order to deliver targeted antibiotic treatments. Toward this end, we present a rapid and integrated single-cell biosensing platform, termed dropFAST, for bacterial growth detection and antimicrobial susceptibility assessment. DropFAST utilizes a rapid resazurin-based fluorescent growth assay coupled with stochastic confinement of bacteria in 20 pL droplets to detect signal from growing bacteria after 1h incubation, equivalent to 2-3 bacterial replications. Full integration of droplet generation, incubation, and detection into a single, uninterrupted stream also renders this platform uniquely suitable for in-line bacterial phenotypic growth assessment. To illustrate the concept of rapid digital antimicrobial susceptibility assessment, we employ the dropFAST platform to evaluate the antibacterial effect of gentamicin on E. coli growth.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close There remains an urgent need for rapid diagnostic methods that can evaluate antibiotic resistance for pathogenic bacteria in order to deliver targeted antibiotic treatments. Toward this end, we present a rapid and integrated single-cell biosensing platform, termed dropFAST, for bacterial growth detection and antimicrobial susceptibility assessment. DropFAST utilizes a rapid resazurin-based fluorescent growth assay coupled with stochastic confinement of bacteria in 20 pL droplets to detect signal from growing bacteria after 1h incubation, equivalent to 2-3 bacterial replications. Full integration of droplet generation, incubation, and detection into a single, uninterrupted stream also renders this platform uniquely suitable for in-line bacterial phenotypic growth assessment. To illustrate the concept of rapid digital antimicrobial susceptibility assessment, we employ the dropFAST platform to evaluate the antibacterial effect of gentamicin on E. coli growth. Close https://www.ncbi.nlm.nih.gov/pubmed/28609716 doi:10.1016/j.bios.2017.06.006 Close
53.	Kilburn, D; Song, Y; Wang, T H; Liu, K J: A Multiplex Ligation Assay for miRNA Copy Number Profiling. In: Methods Mol Biol, 1509 , pp. 185-193, 2017, ISSN: 1940-6029 (Electronic) 1064-3745 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN5269, title = {A Multiplex Ligation Assay for miRNA Copy Number Profiling}, author = { D. Kilburn and Y. Song and T. H. Wang and K. J. Liu}, url = {https://www.ncbi.nlm.nih.gov/pubmed/27826928}, doi = {10.1007/978-1-4939-6524-3_17}, issn = {1940-6029 (Electronic) 1064-3745 (Linking)}, year = {2017}, date = {2017-11-09}, journal = {Methods Mol Biol}, volume = {1509}, pages = {185-193}, abstract = {Ligo-miR is an assay technology that can perform multiplexed detection of miRNAs from a wide range of biological sources. At its core are two sequential ligation steps. First in the capture ligation, template molecules are created by ligating a DNA adapter to the 3' end of all miRNA molecules. Then in the coding ligation these templates are used to generate, linearly amplified, DNA products encoded by length. The resultant number of each DNA product is proportional to the original number of miRNA molecules. The products and their corresponding miRNA can be identified and quantified using common DNA sizing methods such as electrophoresis.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Ligo-miR is an assay technology that can perform multiplexed detection of miRNAs from a wide range of biological sources. At its core are two sequential ligation steps. First in the capture ligation, template molecules are created by ligating a DNA adapter to the 3' end of all miRNA molecules. Then in the coding ligation these templates are used to generate, linearly amplified, DNA products encoded by length. The resultant number of each DNA product is proportional to the original number of miRNA molecules. The products and their corresponding miRNA can be identified and quantified using common DNA sizing methods such as electrophoresis. Close https://www.ncbi.nlm.nih.gov/pubmed/27826928 doi:10.1007/978-1-4939-6524-3_17 Close
54.	Athamanolap, P; Hsieh, K; Chen, L; Yang, S; Wang, T H: Integrated Bacterial Identification and Antimicrobial Susceptibility Testing Using PCR and High-Resolution Melt. In: Anal Chem, 89 (21), pp. 11529-11536, 2017, ISSN: 1520-6882 (Electronic) 0003-2700 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN5648, title = {Integrated Bacterial Identification and Antimicrobial Susceptibility Testing Using PCR and High-Resolution Melt}, author = { P. Athamanolap and K. Hsieh and L. Chen and S. Yang and T. H. Wang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/29027789}, doi = {10.1021/acs.analchem.7b02809}, issn = {1520-6882 (Electronic) 0003-2700 (Linking)}, year = {2017}, date = {2017-11-07}, journal = {Anal Chem}, volume = {89}, number = {21}, pages = {11529-11536}, abstract = {Accurate and timely diagnostics are critical for managing bacterial infections. The current gold standard, culture-based diagnostics, can provide clinicians with comprehensive diagnostic information including bacterial identity and antimicrobial susceptibility, but they often require several days of turnaround time, which leads to compromised clinical outcome and promotes the spread of antibiotic resistance. Nucleic acid amplification tests such as PCR have significantly accelerated the detection of specific bacteria but generally lack the capacities for broad-based bacterial identification or antimicrobial susceptibility testing. Here, we report an integrated assay based on PCR and high-resolution melt (HRM) for rapid diagnosis for bacterial infections. In our assay, we measure bacterial growth in the presence or absence of certain antibiotics with real-time quantitative PCR or digital PCR to determine antimicrobial susceptibility. In addition, we use HRM and a machine learning algorithm to identify bacterial species based on melt-curve profiles of the 16S rRNA gene in an automated fashion. As a demonstration, we correctly identified the bacterial species and their antimicrobial susceptibility profiles for multiple unknown samples in blinded tests within approximately 6.5 h.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Accurate and timely diagnostics are critical for managing bacterial infections. The current gold standard, culture-based diagnostics, can provide clinicians with comprehensive diagnostic information including bacterial identity and antimicrobial susceptibility, but they often require several days of turnaround time, which leads to compromised clinical outcome and promotes the spread of antibiotic resistance. Nucleic acid amplification tests such as PCR have significantly accelerated the detection of specific bacteria but generally lack the capacities for broad-based bacterial identification or antimicrobial susceptibility testing. Here, we report an integrated assay based on PCR and high-resolution melt (HRM) for rapid diagnosis for bacterial infections. In our assay, we measure bacterial growth in the presence or absence of certain antibiotics with real-time quantitative PCR or digital PCR to determine antimicrobial susceptibility. In addition, we use HRM and a machine learning algorithm to identify bacterial species based on melt-curve profiles of the 16S rRNA gene in an automated fashion. As a demonstration, we correctly identified the bacterial species and their antimicrobial susceptibility profiles for multiple unknown samples in blinded tests within approximately 6.5 h. Close https://www.ncbi.nlm.nih.gov/pubmed/29027789 doi:10.1021/acs.analchem.7b02809 Close
55.	Friedrich, S M; Burke, J M; Liu, K J; Ivory, C F; Wang, T H: Molecular rheotaxis directs DNA migration and concentration against a pressure-driven flow. In: Nat Commun, 8 (1), pp. 1213, 2017, ISSN: 2041-1723 (Electronic) 2041-1723 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN5642, title = {Molecular rheotaxis directs DNA migration and concentration against a pressure-driven flow}, author = { S. M. Friedrich and J. M. Burke and K. J. Liu and C. F. Ivory and T. H. Wang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/29089494}, doi = {10.1038/s41467-017-01214-y}, issn = {2041-1723 (Electronic) 2041-1723 (Linking)}, year = {2017}, date = {2017-10-31}, journal = {Nat Commun}, volume = {8}, number = {1}, pages = {1213}, abstract = {In-line preconcentration techniques are used to improve the sensitivity of microfluidic DNA analysis platforms. The most common methods are electrokinetic and require an externally applied electric field. Here we describe a microfluidic DNA preconcentration technique that does not require an external field. Instead, pressure-driven flow from a fluid-filled microcapillary into a lower ionic strength DNA sample reservoir induces spontaneous DNA migration against the direction of flow. This migratory phenomenon that we call Molecular Rheotaxis initiates in seconds and results in a concentrated DNA bolus at the capillary orifice. We demonstrate the ease with which this concentration method can be integrated into a microfluidic total analysis system composed of in-line DNA preconcentration, size separation, and single-molecule detection. Paired experimental and numerical simulation results are used to delineate the parameters required to induce Molecular Rheotaxis, elucidate the underlying mechanism, and optimize conditions to achieve DNA concentration factors exceeding 10,000 fold.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close In-line preconcentration techniques are used to improve the sensitivity of microfluidic DNA analysis platforms. The most common methods are electrokinetic and require an externally applied electric field. Here we describe a microfluidic DNA preconcentration technique that does not require an external field. Instead, pressure-driven flow from a fluid-filled microcapillary into a lower ionic strength DNA sample reservoir induces spontaneous DNA migration against the direction of flow. This migratory phenomenon that we call Molecular Rheotaxis initiates in seconds and results in a concentrated DNA bolus at the capillary orifice. We demonstrate the ease with which this concentration method can be integrated into a microfluidic total analysis system composed of in-line DNA preconcentration, size separation, and single-molecule detection. Paired experimental and numerical simulation results are used to delineate the parameters required to induce Molecular Rheotaxis, elucidate the underlying mechanism, and optimize conditions to achieve DNA concentration factors exceeding 10,000 fold. Close https://www.ncbi.nlm.nih.gov/pubmed/29089494 doi:10.1038/s41467-017-01214-y Close
56.	Axt, B; Hsieh, Y F; Nalayanda, D; Wang, T H: Impedance feedback control of microfluidic valves for reliable post processing combinatorial droplet injection. In: Biomed Microdevices, 19 (3), pp. 61, 2017, ISSN: 1572-8781 (Electronic) 1387-2176 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN5625, title = {Impedance feedback control of microfluidic valves for reliable post processing combinatorial droplet injection}, author = { B. Axt and Y. F. Hsieh and D. Nalayanda and T. H. Wang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/28681238}, doi = {10.1007/s10544-017-0203-2}, issn = {1572-8781 (Electronic) 1387-2176 (Linking)}, year = {2017}, date = {2017-09-01}, journal = {Biomed Microdevices}, volume = {19}, number = {3}, pages = {61}, abstract = {Droplet microfluidics has found use in many biological assay applications as a means of high-throughput sample processing. One of the challenges of the technology, however, is the ability to control and merge droplets on-demand as they flow through the microdevices. It is in the interest of developing lab-on-chip devices to be able to combinatorically program additive mixing steps for more complex multistep and multiplex assays. Existing technologies to merge droplets are either passive in nature or require highly predictable droplet movement for feedforward control, making them vulnerable to errors during high throughput operation. In this paper, we describe and demonstrate a microfluidic valve-based device for the purpose of combinatorial droplet injection at any stage in a multistep assay. Microfluidic valves are used to robustly control fluid flow, droplet generation, and droplet mixing in the device on-demand, while on-chip impedance measurements taken in real time are used as feedback to accurately time the droplet injections. The presented system is contrasted to attempts without feedback, and is shown to be 100% reliable over long durations. Additionally, content detection and discretionary injections are explored and successfully executed.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Droplet microfluidics has found use in many biological assay applications as a means of high-throughput sample processing. One of the challenges of the technology, however, is the ability to control and merge droplets on-demand as they flow through the microdevices. It is in the interest of developing lab-on-chip devices to be able to combinatorically program additive mixing steps for more complex multistep and multiplex assays. Existing technologies to merge droplets are either passive in nature or require highly predictable droplet movement for feedforward control, making them vulnerable to errors during high throughput operation. In this paper, we describe and demonstrate a microfluidic valve-based device for the purpose of combinatorial droplet injection at any stage in a multistep assay. Microfluidic valves are used to robustly control fluid flow, droplet generation, and droplet mixing in the device on-demand, while on-chip impedance measurements taken in real time are used as feedback to accurately time the droplet injections. The presented system is contrasted to attempts without feedback, and is shown to be 100% reliable over long durations. Additionally, content detection and discretionary injections are explored and successfully executed. Close https://www.ncbi.nlm.nih.gov/pubmed/28681238 doi:10.1007/s10544-017-0203-2 Close
57.	Song, Y; Kilburn, D; Song, J H; Cheng, Y; Saeui, C T; Cheung, D G; Croce, C M; Yarema, K J; Meltzer, S J; Liu, K J; Wang, T H: Determination of absolute expression profiles using multiplexed miRNA analysis. In: PLoS One, 12 (7), pp. e0180988, 2017, ISSN: 1932-6203 (Electronic) 1932-6203 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN5621, title = {Determination of absolute expression profiles using multiplexed miRNA analysis}, author = { Y. Song and D. Kilburn and J. H. Song and Y. Cheng and C. T. Saeui and D. G. Cheung and C. M. Croce and K. J. Yarema and S. J. Meltzer and K. J. Liu and T. H. Wang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/28704432}, doi = {10.1371/journal.pone.0180988}, issn = {1932-6203 (Electronic) 1932-6203 (Linking)}, year = {2017}, date = {2017-07-13}, journal = {PLoS One}, volume = {12}, number = {7}, pages = {e0180988}, abstract = {Accurate measurement of miRNA expression is critical to understanding their role in gene expression as well as their application as disease biomarkers. Correct identification of changes in miRNA expression rests on reliable normalization to account for biological and technological variance between samples. Ligo-miR is a multiplex assay designed to rapidly measure absolute miRNA copy numbers, thus reducing dependence on biological controls. It uses a simple 2-step ligation process to generate length coded products that can be quantified using a variety of DNA sizing methods. We demonstrate Ligo-miR's ability to quantify miRNA expression down to 20 copies per cell sensitivity, accurately discriminate between closely related miRNA, and reliably measure differential changes as small as 1.2-fold. Then, benchmarking studies were performed to show the high correlation between Ligo-miR, microarray, and TaqMan qRT-PCR. Finally, Ligo-miR was used to determine copy number profiles in a number of breast, esophageal, and pancreatic cell lines and to demonstrate the utility of copy number analysis for providing layered insight into expression profile changes.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Accurate measurement of miRNA expression is critical to understanding their role in gene expression as well as their application as disease biomarkers. Correct identification of changes in miRNA expression rests on reliable normalization to account for biological and technological variance between samples. Ligo-miR is a multiplex assay designed to rapidly measure absolute miRNA copy numbers, thus reducing dependence on biological controls. It uses a simple 2-step ligation process to generate length coded products that can be quantified using a variety of DNA sizing methods. We demonstrate Ligo-miR's ability to quantify miRNA expression down to 20 copies per cell sensitivity, accurately discriminate between closely related miRNA, and reliably measure differential changes as small as 1.2-fold. Then, benchmarking studies were performed to show the high correlation between Ligo-miR, microarray, and TaqMan qRT-PCR. Finally, Ligo-miR was used to determine copy number profiles in a number of breast, esophageal, and pancreatic cell lines and to demonstrate the utility of copy number analysis for providing layered insight into expression profile changes. Close https://www.ncbi.nlm.nih.gov/pubmed/28704432 doi:10.1371/journal.pone.0180988 Close
58.	Shin, D J; Athamanolap, P; Chen, L; Hardick, J; Lewis, M; Hsieh, Y H; Rothman, R E; Gaydos, C A; Wang, T H: Mobile nucleic acid amplification testing (mobiNAAT) for Chlamydia trachomatis screening in hospital emergency department settings. In: Sci Rep, 7 (1), pp. 4495, 2017, ISSN: 2045-2322 (Electronic) 2045-2322 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN5632, title = {Mobile nucleic acid amplification testing (mobiNAAT) for Chlamydia trachomatis screening in hospital emergency department settings}, author = { D. J. Shin and P. Athamanolap and L. Chen and J. Hardick and M. Lewis and Y. H. Hsieh and R. E. Rothman and C. A. Gaydos and T. H. Wang}, url = {https://www.ncbi.nlm.nih.gov/pubmed/28674410}, doi = {10.1038/s41598-017-04781-8}, issn = {2045-2322 (Electronic) 2045-2322 (Linking)}, year = {2017}, date = {2017-07-03}, journal = {Sci Rep}, volume = {7}, number = {1}, pages = {4495}, abstract = {Management of curable sexually-transmitted infections (STI) such as Chlamydia can be revolutionized by highly sensitive nucleic acid testing that is deployable at the point-of-care (POC). Here we report the development of a mobile nucleic acid amplification testing (mobiNAAT) platform utilizing a mobile phone and droplet magnetofluidics to deliver NAAT in a portable and accessible format. By using magnetic particles as a mobile substrate for nucleic acid capture and transport, fluid handling is reduced to particle translocation on a simple magnetofluidic cartridge assembled with reagents for nucleic acid purification and amplification. A mobile phone user interface operating in tandem with a portable Bluetooth-enabled cartridge-processing unit facilitates process integration. We tested 30 potentially Chlamydia trachomatis (CT)-infected patients in a hospital emergency department and confirmed that mobiNAAT showed 100% concordance with laboratory-based NAAT. Concurrent evaluation by a nontechnical study coordinator who received brief training via an embedded mobile app module demonstrated ease of use and reproducibility of the platform. This work demonstrates the potential of mobile nucleic acid testing in bridging the diagnostic gap between centralized laboratories and hospital emergency departments.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Management of curable sexually-transmitted infections (STI) such as Chlamydia can be revolutionized by highly sensitive nucleic acid testing that is deployable at the point-of-care (POC). Here we report the development of a mobile nucleic acid amplification testing (mobiNAAT) platform utilizing a mobile phone and droplet magnetofluidics to deliver NAAT in a portable and accessible format. By using magnetic particles as a mobile substrate for nucleic acid capture and transport, fluid handling is reduced to particle translocation on a simple magnetofluidic cartridge assembled with reagents for nucleic acid purification and amplification. A mobile phone user interface operating in tandem with a portable Bluetooth-enabled cartridge-processing unit facilitates process integration. We tested 30 potentially Chlamydia trachomatis (CT)-infected patients in a hospital emergency department and confirmed that mobiNAAT showed 100% concordance with laboratory-based NAAT. Concurrent evaluation by a nontechnical study coordinator who received brief training via an embedded mobile app module demonstrated ease of use and reproducibility of the platform. This work demonstrates the potential of mobile nucleic acid testing in bridging the diagnostic gap between centralized laboratories and hospital emergency departments. Close https://www.ncbi.nlm.nih.gov/pubmed/28674410 doi:10.1038/s41598-017-04781-8 Close
59.	Davenport, M; Mach, K E; Shortliffe, L M D; Banaei, N; Wang, T H; Liao, J C: New and developing diagnostic technologies for urinary tract infections. In: Nat Rev Urol, 14 (5), pp. 296-310, 2017, ISSN: 1759-4820 (Electronic) 1759-4812 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN5273, title = {New and developing diagnostic technologies for urinary tract infections}, author = { M. Davenport and K. E. Mach and L. M. D. Shortliffe and N. Banaei and T. H. Wang and J. C. Liao}, url = {https://www.ncbi.nlm.nih.gov/pubmed/28248946}, doi = {10.1038/nrurol.2017.20}, issn = {1759-4820 (Electronic) 1759-4812 (Linking)}, year = {2017}, date = {2017-05-01}, journal = {Nat Rev Urol}, volume = {14}, number = {5}, pages = {296-310}, abstract = {Timely and accurate identification and determination of the antimicrobial susceptibility of uropathogens is central to the management of UTIs. Urine dipsticks are fast and amenable to point-of-care testing, but do not have adequate diagnostic accuracy or provide microbiological diagnosis. Urine culture with antimicrobial susceptibility testing takes 2-3 days and requires a clinical laboratory. The common use of empirical antibiotics has contributed to the rise of multidrug-resistant organisms, reducing treatment options and increasing costs. In addition to improved antimicrobial stewardship and the development of new antimicrobials, novel diagnostics are needed for timely microbial identification and determination of antimicrobial susceptibilities. New diagnostic platforms, including nucleic acid tests and mass spectrometry, have been approved for clinical use and have improved the speed and accuracy of pathogen identification from primary cultures. Optimization for direct urine testing would reduce the time to diagnosis, yet these technologies do not provide comprehensive information on antimicrobial susceptibility. Emerging technologies including biosensors, microfluidics, and other integrated platforms could improve UTI diagnosis via direct pathogen detection from urine samples, rapid antimicrobial susceptibility testing, and point-of-care testing. Successful development and implementation of these technologies has the potential to usher in an era of precision medicine to improve patient care and public health.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Timely and accurate identification and determination of the antimicrobial susceptibility of uropathogens is central to the management of UTIs. Urine dipsticks are fast and amenable to point-of-care testing, but do not have adequate diagnostic accuracy or provide microbiological diagnosis. Urine culture with antimicrobial susceptibility testing takes 2-3 days and requires a clinical laboratory. The common use of empirical antibiotics has contributed to the rise of multidrug-resistant organisms, reducing treatment options and increasing costs. In addition to improved antimicrobial stewardship and the development of new antimicrobials, novel diagnostics are needed for timely microbial identification and determination of antimicrobial susceptibilities. New diagnostic platforms, including nucleic acid tests and mass spectrometry, have been approved for clinical use and have improved the speed and accuracy of pathogen identification from primary cultures. Optimization for direct urine testing would reduce the time to diagnosis, yet these technologies do not provide comprehensive information on antimicrobial susceptibility. Emerging technologies including biosensors, microfluidics, and other integrated platforms could improve UTI diagnosis via direct pathogen detection from urine samples, rapid antimicrobial susceptibility testing, and point-of-care testing. Successful development and implementation of these technologies has the potential to usher in an era of precision medicine to improve patient care and public health. Close https://www.ncbi.nlm.nih.gov/pubmed/28248946 doi:10.1038/nrurol.2017.20 Close
60.	Hulbert, A; Jusue-Torres, I; Stark, A; Chen, C; Rodgers, K; Lee, B; Griffin, C; Yang, A; Huang, P; Wrangle, J; Belinsky, S A; Wang, T H; Yang, S C; Baylin, S B; Brock, M V; Herman, J G: Early Detection of Lung Cancer Using DNA Promoter Hypermethylation in Plasma and Sputum. In: Clin Cancer Res, 23 (8), pp. 1998-2005, 2017, ISSN: 1078-0432 (Print) 1078-0432 (Linking). (Type: Journal Article \| Abstract \| Links \| BibTeX) @article{RN5160, title = {Early Detection of Lung Cancer Using DNA Promoter Hypermethylation in Plasma and Sputum}, author = { A. Hulbert and I. Jusue-Torres and A. Stark and C. Chen and K. Rodgers and B. Lee and C. Griffin and A. Yang and P. Huang and J. Wrangle and S. A. Belinsky and T. H. Wang and S. C. Yang and S. B. Baylin and M. V. Brock and J. G. Herman}, url = {https://www.ncbi.nlm.nih.gov/pubmed/27729459}, doi = {10.1158/1078-0432.CCR-16-1371}, issn = {1078-0432 (Print) 1078-0432 (Linking)}, year = {2017}, date = {2017-04-15}, journal = {Clin Cancer Res}, volume = {23}, number = {8}, pages = {1998-2005}, abstract = {Purpose: CT screening can reduce death from lung cancer. We sought to improve the diagnostic accuracy of lung cancer screening using ultrasensitive methods and a lung cancer-specific gene panel to detect DNA methylation in sputum and plasma.Experimental Design: This is a case-control study of subjects with suspicious nodules on CT imaging. Plasma and sputum were obtained preoperatively. Cases (n = 150) had pathologic confirmation of node-negative (stages I and IIA) non-small cell lung cancer. Controls (n = 60) had non-cancer diagnoses. We detected promoter methylation using quantitative methylation-specific real-time PCR and methylation-on-beads for cancer-specific genes (SOX17, TAC1, HOXA7, CDO1, HOXA9, and ZFP42).Results: DNA methylation was detected in plasma and sputum more frequently in people with cancer compared with controls (P < 0.001) for five of six genes. The sensitivity and specificity for lung cancer diagnosis using the best individual genes was 63% to 86% and 75% to 92% in sputum, respectively, and 65% to 76% and 74% to 84% in plasma, respectively. A three-gene combination of the best individual genes has sensitivity and specificity of 98% and 71% using sputum and 93% and 62% using plasma. Area under the receiver operating curve for this panel was 0.89 [95% confidence interval (CI), 0.80-0.98] in sputum and 0.77 (95% CI, 0.68-0.86) in plasma. Independent blinded random forest prediction models combining gene methylation with clinical information correctly predicted lung cancer in 91% of subjects using sputum detection and 85% of subjects using plasma detection.Conclusions: High diagnostic accuracy for early-stage lung cancer can be obtained using methylated promoter detection in sputum or plasma. Clin Cancer Res; 23(8); 1998-2005. (c)2016 AACR.}, keywords = {}, pubstate = {published}, tppubtype = {article} } Close Purpose: CT screening can reduce death from lung cancer. We sought to improve the diagnostic accuracy of lung cancer screening using ultrasensitive methods and a lung cancer-specific gene panel to detect DNA methylation in sputum and plasma.Experimental Design: This is a case-control study of subjects with suspicious nodules on CT imaging. Plasma and sputum were obtained preoperatively. Cases (n = 150) had pathologic confirmation of node-negative (stages I and IIA) non-small cell lung cancer. Controls (n = 60) had non-cancer diagnoses. We detected promoter methylation using quantitative methylation-specific real-time PCR and methylation-on-beads for cancer-specific genes (SOX17, TAC1, HOXA7, CDO1, HOXA9, and ZFP42).Results: DNA methylation was detected in plasma and sputum more frequently in people with cancer compared with controls (P < 0.001) for five of six genes. The sensitivity and specificity for lung cancer diagnosis using the best individual genes was 63% to 86% and 75% to 92% in sputum, respectively, and 65% to 76% and 74% to 84% in plasma, respectively. A three-gene combination of the best individual genes has sensitivity and specificity of 98% and 71% using sputum and 93% and 62% using plasma. Area under the receiver operating curve for this panel was 0.89 [95% confidence interval (CI), 0.80-0.98] in sputum and 0.77 (95% CI, 0.68-0.86) in plasma. Independent blinded random forest prediction models combining gene methylation with clinical information correctly predicted lung cancer in 91% of subjects using sputum detection and 85% of subjects using plasma detection.Conclusions: High diagnostic accuracy for early-stage lung cancer can be obtained using methylated promoter detection in sputum or plasma. Clin Cancer Res; 23(8); 1998-2005. (c)2016 AACR. Close https://www.ncbi.nlm.nih.gov/pubmed/27729459 doi:10.1158/1078-0432.CCR-16-1371 Close

152 entries « ‹ 3 of 8 › »

152 entries « ‹ 3 of 4 › »

2014

D. J. Shin; T. H. Wang

Magnetic droplet manipulation platforms for nucleic acid detection at the point of care Journal Article

Ann Biomed Eng, 42 (11), pp. 2289-302, 2014, ISSN: 1573-9686 (Electronic) 0090-6964 (Linking).

Abstract | Links | BibTeX

P. Athamanolap; V. Parekh; S. I. Fraley; V. Agarwal; D. J. Shin; M. A. Jacobs; T. H. Wang; S. Yang

Trainable high resolution melt curve machine learning classifier for large-scale reliable genotyping of sequence variants Journal Article

PLoS One, 9 (9), pp. e109094, 2014, ISSN: 1932-6203 (Electronic) 1932-6203 (Linking).

Abstract | Links | BibTeX

A. A. Guzzetta; T. R. Pisanic Ii; P. Sharma; J. M. Yi; A. Stark; T. H. Wang; N. Ahuja

The promise of methylation on beads for cancer detection and treatment Journal Article

Expert Rev Mol Diagn, 14 (7), pp. 845-52, 2014, ISSN: 1744-8352 (Electronic) 1473-7159 (Linking).

Abstract | Links | BibTeX

H. Zec; D. J. Shin; T. H. Wang

Novel droplet platforms for the detection of disease biomarkers Journal Article

Expert Rev Mol Diagn, 14 (7), pp. 787-801, 2014, ISSN: 1744-8352 (Electronic) 1473-7159 (Linking).

Abstract | Links | BibTeX

C. W. Beh; D. Pan; J. Lee; X. Jiang; K. J. Liu; H. Q. Mao; T. H. Wang

Direct interrogation of DNA content distribution in nanoparticles by a novel microfluidics-based single-particle analysis Journal Article

Nano Lett, 14 (8), pp. 4729-35, 2014, ISSN: 1530-6992 (Electronic) 1530-6984 (Linking).

Abstract | Links | BibTeX

@article{RN3208,
title = {Direct interrogation of DNA content distribution in nanoparticles by a novel microfluidics-based single-particle analysis},
author = { C. W. Beh and D. Pan and J. Lee and X. Jiang and K. J. Liu and H. Q. Mao and T. H. Wang},
url = {https://www.ncbi.nlm.nih.gov/pubmed/25054542},
doi = {10.1021/nl5018404},
issn = {1530-6992 (Electronic)
1530-6984 (Linking)},
year = {2014},
date = {2014-08-13},
journal = {Nano Lett},
volume = {14},
number = {8},
pages = {4729-35},
abstract = {Nonviral gene delivery holds great promise not just as a safer alternative to viral vectors in traditional gene therapy applications, but also for regenerative medicine, induction of pluripotency in somatic cells, and RNA interference for gene silencing. Although it continues to be an active area of research, there remain many challenges to the rational design of vectors. Among these, the inability to characterize the composition of nanoparticles and its distribution has made it difficult to probe the mechanism of gene transfection process, since differences in the nanoparticle-mediated transfection exist even when the same vector is used. There is a lack of sensitive methods that allow for full characterization of DNA content in single nanoparticles and its distribution among particles in the same preparation. Here we report a novel spectroscopic approach that is capable of interrogating nanoparticles on a particle-by-particle basis. Using PEI/DNA and PEI-g-PEG/DNA nanoparticles as examples, we have shown that the distribution of DNA content among these nanoparticles was relatively narrow, with the average numbers of DNA of 4.8 and 6.7 per particle, respectively, in PEI/DNA and PEI-g-PEG/DNA nanoparticles. This analysis enables a more accurate description of DNA content in polycation/DNA nanoparticles. It paves the way toward comparative assessments of various types of gene carriers and provides insights into bridging the efficiency gap between viral and nonviral vehicles.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}

D. J. Shin; Y. Zhang; T. H. Wang

A droplet microfluidic approach to single-stream nucleic acid isolation and mutation detection Journal Article

Microfluid Nanofluidics, 17 (2), pp. 425-430, 2014, ISSN: 1613-4982 (Print) 1613-4982 (Linking).

Abstract | Links | BibTeX

T. R. 2nd Pisanic; Y. Zhang; T. H. Wang

Quantum dots in diagnostics and detection: principles and paradigms Journal Article

Analyst, 139 (12), pp. 2968-81, 2014, ISSN: 1364-5528 (Electronic) 0003-2654 (Linking).

Abstract | Links | BibTeX

P. Athamanolap; D. J. Shin; T. H. Wang

Droplet Array Platform for High-Resolution Melt Analysis of DNA Methylation Density Journal Article

J Lab Autom, 19 (3), pp. 304-12, 2014, ISSN: 2211-0690 (Electronic) 2211-0682 (Linking).

Abstract | Links | BibTeX

Y. Song; K. J. Liu; T. H. Wang

Elimination of ligation dependent artifacts in T4 RNA ligase to achieve high efficiency and low bias microRNA capture Journal Article

PLoS One, 9 (4), pp. e94619, 2014, ISSN: 1932-6203 (Electronic) 1932-6203 (Linking).

Abstract | Links | BibTeX

L. Zhang; Y. Song; T. Fujita; Y. Zhang; M. Chen; T. H. Wang

Large enhancement of quantum dot fluorescence by highly scalable nanoporous gold Journal Article

Adv Mater, 26 (8), pp. 1289-94, 2014, ISSN: 1521-4095 (Electronic) 0935-9648 (Linking).

Abstract | Links | BibTeX

D. D. Nalayanda; W. B. Fulton; P. M. Colombani; T. H. Wang; F. Abdullah

Pressure induced lung injury in a novel in vitro model of the alveolar interface: protective effect of dexamethasone Journal Article

J Pediatr Surg, 49 (1), pp. 61-5; discussion 65, 2014, ISSN: 1531-5037 (Electronic) 0022-3468 (Linking).

Abstract | Links | BibTeX

@article{RN3212,
title = {Pressure induced lung injury in a novel in vitro model of the alveolar interface: protective effect of dexamethasone},
author = { D. D. Nalayanda and W. B. Fulton and P. M. Colombani and T. H. Wang and F. Abdullah},
url = {https://www.ncbi.nlm.nih.gov/pubmed/24439582},
doi = {10.1016/j.jpedsurg.2013.09.030},
issn = {1531-5037 (Electronic)
0022-3468 (Linking)},
year = {2014},
date = {2014-01-01},
journal = {J Pediatr Surg},
volume = {49},
number = {1},
pages = {61-5; discussion 65},
abstract = {PURPOSE: The lungs of infants born with congenital diaphragmatic hernia suffer from immaturity as well as the short and long term consequences of ventilator-induced lung injury, including chronic lung disease. Antenatal and postnatal steroids are among current strategies promoted to treat premature lungs and limit long term morbidity. Although studied in whole-animal models, insight into ventilator-induced injury at the alveolar-capillary interface as well as the benefits of steroids, remains limited. The present study utilizes a multi-fluidic in vitro model of the alveolar-interface to analyze membrane disruption from compressive aerodynamic forces in dexamethasone-treated cultures. METHODS: Human alveolar epithelial cell lines, H441 and A549, were cultured in a custom-built chamber under constant aerodynamic shear followed by introduction of pressure stimuli with and without dexamethasone (0.1muM). On-chip bioelectrical measurements were noted to track changes to the cellular surface and live-dead assay to ascertain cellular viability. RESULTS: Pressure-exposed alveolar cultures demonstrated a significant drop in TEER that was less prominent with an underlying extracellular-matrix coating. Addition of dexamethasone resulted in increased alveolar layer integrity demonstrated by higher TEER values. Furthermore, dexamethasone-treated cells exhibited faster recovery, and the effects of pressure appeared to be mitigated in both cell types. CONCLUSION: Using a novel in vitro model of the alveolus, we demonstrate a dose-response relationship between pressure application and loss of alveolar layer integrity. This effect appears to be alleviated by dexamethasone and matrix sub-coating.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}

2013

Y. Zhang; D. J. Shin; T. H. Wang

Serial dilution via surface energy trap-assisted magnetic droplet manipulation Journal Article

Lab Chip, 13 (24), pp. 4827-31, 2013, ISSN: 1473-0189 (Electronic) 1473-0189 (Linking).

Abstract | Links | BibTeX

C. H. Chiou; D. J. Shin; Y. Zhang; T. H. Wang

Topography-assisted electromagnetic platform for blood-to-PCR in a droplet Journal Article

Biosens Bioelectron, 50 , pp. 91-9, 2013, ISSN: 1873-4235 (Electronic) 0956-5663 (Linking).

Abstract | Links | BibTeX

J. M. Yi; A. A. Guzzetta; V. J. Bailey; S. R. Downing; L. Van Neste; K. B. Chiappinelli; B. P. Keeley; A. Stark; A. Herrera; C. Wolfgang; E. P. Pappou; C. A. Iacobuzio-Donahue; M. G. Goggins; J. G. Herman; T. H. Wang; S. B. Baylin; N. Ahuja

Novel methylation biomarker panel for the early detection of pancreatic cancer Journal Article

Clin Cancer Res, 19 (23), pp. 6544-6555, 2013, ISSN: 1078-0432 (Print) 1078-0432 (Linking).

Abstract | Links | BibTeX

@article{RN3216,
title = {Novel methylation biomarker panel for the early detection of pancreatic cancer},
author = { J. M. Yi and A. A. Guzzetta and V. J. Bailey and S. R. Downing and L. Van Neste and K. B. Chiappinelli and B. P. Keeley and A. Stark and A. Herrera and C. Wolfgang and E. P. Pappou and C. A. Iacobuzio-Donahue and M. G. Goggins and J. G. Herman and T. H. Wang and S. B. Baylin and N. Ahuja},
url = {https://www.ncbi.nlm.nih.gov/pubmed/24088737},
doi = {10.1158/1078-0432.CCR-12-3224},
issn = {1078-0432 (Print)
1078-0432 (Linking)},
year = {2013},
date = {2013-12-01},
journal = {Clin Cancer Res},
volume = {19},
number = {23},
pages = {6544-6555},
abstract = {PURPOSE: Pancreatic cancer is the fourth leading cause of cancer deaths and there currently is no reliable modality for the early detection of this disease. Here, we identify cancer-specific promoter DNA methylation of BNC1 and ADAMTS1 as a promising biomarker detection strategy meriting investigation in pancreatic cancer. EXPERIMENTAL DESIGN: We used a genome-wide pharmacologic transcriptome approach to identify novel cancer-specific DNA methylation alterations in pancreatic cancer cell lines. Of eight promising genes, we focused our studies on BNC1 and ADAMTS1 for further downstream analysis, including methylation and expression. We used a nanoparticle-enabled methylation on beads (MOB) technology to detect early-stage pancreatic cancers by analyzing DNA methylation in patient serum. RESULTS: We identified two novel genes, BNC1 (92%) and ADAMTS1 (68%), that showed a high frequency of methylation in pancreatic cancers (n = 143), up to 100% in PanIN-3 and 97% in stage I invasive cancers. Using the nanoparticle-enabled MOB technology, these alterations could be detected in serum samples (n = 42) from patients with pancreatic cancer, with a sensitivity for BNC1 of 79% [95% confidence interval (CI), 66%-91%] and for ADAMTS1 of 48% (95% CI, 33%-63%), whereas specificity was 89% for BNC1 (95% CI, 76%-100%) and 92% for ADAMTS1 (95% CI, 82%-100%). Overall sensitivity using both markers is 81% (95% CI, 69%-93%) and specificity is 85% (95% CI, 71%-99%). CONCLUSIONS: Promoter DNA methylation of BNC1 and ADAMTS1 is a potential biomarker to detect early-stage pancreatic cancers. Assaying the promoter methylation status of these genes in circulating DNA from serum is a promising strategy for early detection of pancreatic cancer and has the potential to improve mortality from this disease.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}

B. Keeley; A. Stark; T. R. 2nd Pisanic; R. Kwak; Y. Zhang; J. Wrangle; S. Baylin; J. Herman; N. Ahuja; M. V. Brock; T. H. Wang

Extraction and processing of circulating DNA from large sample volumes using methylation on beads for the detection of rare epigenetic events Journal Article

Clin Chim Acta, 425 , pp. 169-75, 2013, ISSN: 1873-3492 (Electronic) 0009-8981 (Linking).

Abstract | Links | BibTeX

S. I. Fraley; J. Hardick; B. J. Masek; P. Athamanolap; R. E. Rothman; C. A. Gaydos; K. C. Carroll; T. Wakefield; T. H. Wang; S. Yang

Universal digital high-resolution melt: a novel approach to broad-based profiling of heterogeneous biological samples Journal Article

Nucleic Acids Res, 41 (18), pp. e175, 2013, ISSN: 1362-4962 (Electronic) 0305-1048 (Linking).

Abstract | Links | BibTeX

@article{RN3217,
title = {Universal digital high-resolution melt: a novel approach to broad-based profiling of heterogeneous biological samples},
author = { S. I. Fraley and J. Hardick and B. J. Masek and P. Athamanolap and R. E. Rothman and C. A. Gaydos and K. C. Carroll and T. Wakefield and T. H. Wang and S. Yang},
url = {https://www.ncbi.nlm.nih.gov/pubmed/23935121},
doi = {10.1093/nar/gkt684},
issn = {1362-4962 (Electronic)
0305-1048 (Linking)},
year = {2013},
date = {2013-10-01},
journal = {Nucleic Acids Res},
volume = {41},
number = {18},
pages = {e175},
abstract = {Comprehensive profiling of nucleic acids in genetically heterogeneous samples is important for clinical and basic research applications. Universal digital high-resolution melt (U-dHRM) is a new approach to broad-based PCR diagnostics and profiling technologies that can overcome issues of poor sensitivity due to contaminating nucleic acids and poor specificity due to primer or probe hybridization inaccuracies for single nucleotide variations. The U-dHRM approach uses broad-based primers or ligated adapter sequences to universally amplify all nucleic acid molecules in a heterogeneous sample, which have been partitioned, as in digital PCR. Extensive assay optimization enables direct sequence identification by algorithm-based matching of melt curve shape and Tm to a database of known sequence-specific melt curves. We show that single-molecule detection and single nucleotide sensitivity is possible. The feasibility and utility of U-dHRM is demonstrated through detection of bacteria associated with polymicrobial blood infection and microRNAs (miRNAs) associated with host response to infection. U-dHRM using broad-based 16S rRNA gene primers demonstrates universal single cell detection of bacterial pathogens, even in the presence of larger amounts of contaminating bacteria; U-dHRM using universally adapted Lethal-7 miRNAs in a heterogeneous mixture showcases the single copy sensitivity and single nucleotide specificity of this approach.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}

D. D. Nalayanda; W. B. Fulton; T. H. Wang; F. Abdullah

A multiphase fluidic platform for studying ventilator-induced injury of the pulmonary epithelial barrier Journal Article

Integr Biol (Camb), 5 (9), pp. 1141-8, 2013, ISSN: 1757-9708 (Electronic) 1757-9694 (Linking).

Abstract | Links | BibTeX

T. H. Wang

Micro and nanotechnologies enhanced biomolecular sensing Journal Article

Biosensors (Basel), 3 (3), pp. 283-5, 2013, ISSN: 2079-6374 (Print) 2079-6374 (Linking).

Abstract | Links | BibTeX

Y. Zhang; T. H. Wang

Full-range magnetic manipulation of droplets via surface energy traps enables complex bioassays Journal Article

Adv Mater, 25 (21), pp. 2903-8, 2013, ISSN: 1521-4095 (Electronic) 0935-9648 (Linking).

Abstract | Links | BibTeX

Y. Song; Y. Zhang; T. H. Wang

Single quantum dot analysis enables multiplexed point mutation detection by gap ligase chain reaction Journal Article

Small, 9 (7), pp. 1096-105, 2013, ISSN: 1613-6829 (Electronic) 1613-6810 (Linking).

Abstract | Links | BibTeX

T. H. Wang

Discerning single molecule interactions of DNA and quantum dots Journal Article

Biotechnol J, 8 (1), pp. 15-6, 2013, ISSN: 1860-7314 (Electronic) 1860-6768 (Linking).

Abstract | Links | BibTeX

2012

C. W. Beh; W. Zhou; T. H. Wang

PDMS-glass bonding using grafted polymeric adhesive--alternative process flow for compatibility with patterned biological molecules Journal Article

Lab Chip, 12 (20), pp. 4120-7, 2012, ISSN: 1473-0189 (Electronic) 1473-0189 (Linking).

Abstract | Links | BibTeX

T. D. Rane; H. C. Zec; T. H. Wang

A serial sample loading system: interfacing multiwell plates with microfluidic devices Journal Article

J Lab Autom, 17 (5), pp. 370-7, 2012, ISSN: 2211-0690 (Electronic) 2211-0682 (Linking).

Abstract | Links | BibTeX

T. D. Rane; H. C. Zec; C. Puleo; A. P. Lee; T. H. Wang

Droplet microfluidics for amplification-free genetic detection of single cells Journal Article

Lab Chip, 12 (18), pp. 3341-7, 2012, ISSN: 1473-0189 (Electronic) 1473-0189 (Linking).

Abstract | Links | BibTeX

H. Zec; T. D. Rane; T. H. Wang

Microfluidic platform for on-demand generation of spatially indexed combinatorial droplets Journal Article

Lab Chip, 12 (17), pp. 3055-62, 2012, ISSN: 1473-0189 (Electronic) 1473-0189 (Linking).

Abstract | Links | BibTeX

Y. Zhang; T. H. Wang

Quantum dot enabled molecular sensing and diagnostics Journal Article

Theranostics, 2 (7), pp. 631-54, 2012, ISSN: 1838-7640 (Electronic) 1838-7640 (Linking).

Abstract | Links | BibTeX

@article{RN3225,
title = {Quantum dot enabled molecular sensing and diagnostics},
author = { Y. Zhang and T. H. Wang},
url = {https://www.ncbi.nlm.nih.gov/pubmed/22916072},
doi = {10.7150/thno.4308},
issn = {1838-7640 (Electronic)
1838-7640 (Linking)},
year = {2012},
date = {2012-08-24},
journal = {Theranostics},
volume = {2},
number = {7},
pages = {631-54},
abstract = {Since its emergence, semiconductor nanoparticles known as quantum dots (QDs) have drawn considerable attention and have quickly extended their applicability to numerous fields within the life sciences. This is largely due to their unique optical properties such as high brightness and narrow emission band as well as other advantages over traditional organic fluorophores. New molecular sensing strategies based on QDs have been developed in pursuit of high sensitivity, high throughput, and multiplexing capabilities. For traditional biological applications, QDs have already begun to replace traditional organic fluorophores to serve as simple fluorescent reporters in immunoassays, microarrays, fluorescent imaging applications, and other assay platforms. In addition, smarter, more advanced QD probes such as quantum dot fluorescence resonance energy transfer (QD-FRET) sensors, quenching sensors, and barcoding systems are paving the way for highly-sensitive genetic and epigenetic detection of diseases, multiplexed identification of infectious pathogens, and tracking of intracellular drug and gene delivery. When combined with microfluidics and confocal fluorescence spectroscopy, the detection limit is further enhanced to single molecule level. Recently, investigations have revealed that QDs participate in series of new phenomena and exhibit interesting non-photoluminescent properties. Some of these new findings are now being incorporated into novel assays for gene copy number variation (CNV) studies and DNA methylation analysis with improved quantification resolution. Herein, we provide a comprehensive review on the latest developments of QD based molecular diagnostic platforms in which QD plays a versatile and essential role.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}

Y. Zhang; T. H. Wang

Micro Magnetic Gyromixer for Speeding up Reactions in Droplets Journal Article

Microfluid Nanofluidics, 12 (5), pp. 787-794, 2012, ISSN: 1613-4982 (Print) 1613-4982 (Linking).

Abstract | Links | BibTeX

Y. Zhang; K. J. Liu; T. L. Wang; M. Shih Ie; T. H. Wang

Mapping DNA quantity into electrophoretic mobility through quantum dot nanotethers for high-resolution genetic and epigenetic analysis Journal Article

ACS Nano, 6 (1), pp. 858-64, 2012, ISSN: 1936-086X (Electronic) 1936-0851 (Linking).

Abstract | Links | BibTeX

@article{RN3231,
title = {Mapping DNA quantity into electrophoretic mobility through quantum dot nanotethers for high-resolution genetic and epigenetic analysis},
author = { Y. Zhang and K. J. Liu and T. L. Wang and M. Shih Ie and T. H. Wang},
url = {https://www.ncbi.nlm.nih.gov/pubmed/22136600},
doi = {10.1021/nn204377k},
issn = {1936-086X (Electronic)
1936-0851 (Linking)},
year = {2012},
date = {2012-01-24},
journal = {ACS Nano},
volume = {6},
number = {1},
pages = {858-64},
abstract = {Newly discovered nanoparticle properties have driven the development of novel applications and uses. We report a new observation where the electrophoretic mobility of a quantum dot/DNA nanoassembly can be precisely modulated by the degree of surface DNA conjugation. By using streptavidin-coated quantum dots (QDs) as nanotethers to gather biotin-labeled DNA into electrophoretic nanoassemblies, the QD surface charge is modulated and transformed into electrophoretic mobility shifts using standard agarose gel electrophoresis. Typical fluorescent assays quantify based on relative intensity. However, this phenomenon uses a novel approach that accurately maps DNA quantity into shifts in relative band position. This property was applied in a QD-enabled nanoassay called quantum dot electrophoretic mobility shift assay (QEMSA) that enables accurate quantification of DNA targets down to 1.1-fold (9%) changes in quantity, beyond what is achievable in qPCR. In addition to these experimental findings, an analytical model is presented to explain this behavior. Finally, QEMSA was applied to both genetic and epigenetic analysis of cancer. First, it was used to analyze copy number variation (CNV) of the RSF1/HBXAP gene, where conventional approaches for CNV analysis based on comparative genomic hybridization (CGH), microarrays, and qPCR are unable to reliably differentiate less than 2-fold changes in copy number. Then, QEMSA was used for DNA methylation analysis of the p16/CDK2A tumor suppressor gene, where its ability to detect subtle changes in methylation was shown to be superior to that of qPCR.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}

2011

S. Park; Y. Zhang; S. Lin; T. H. Wang; S. Yang

Advances in microfluidic PCR for point-of-care infectious disease diagnostics Journal Article

Biotechnol Adv, 29 (6), pp. 830-9, 2011, ISSN: 1873-1899 (Electronic) 0734-9750 (Linking).

Abstract | Links | BibTeX

S. Park; Y. Zhang; T. H. Wang; S. Yang

Continuous dielectrophoretic bacterial separation and concentration from physiological media of high conductivity Journal Article

Lab Chip, 11 (17), pp. 2893-900, 2011, ISSN: 1473-0189 (Electronic) 1473-0189 (Linking).

Abstract | Links | BibTeX

@article{RN3232,
title = {Continuous dielectrophoretic bacterial separation and concentration from physiological media of high conductivity},
author = { S. Park and Y. Zhang and T. H. Wang and S. Yang},
url = {https://www.ncbi.nlm.nih.gov/pubmed/21776517},
doi = {10.1039/c1lc20307j},
issn = {1473-0189 (Electronic)
1473-0189 (Linking)},
year = {2011},
date = {2011-09-07},
journal = {Lab Chip},
volume = {11},
number = {17},
pages = {2893-900},
abstract = {Biological sample processing involves purifying target analytes from various sample matrices and concentrating them to a small volume from a large volume of crude sample. This complex process is the major obstacle for developing a microfluidic diagnostic platform. In this study, we present a microfluidic device that can continuously separate and concentrate pathogenic bacterial cells from complex sample matrices such as cerebrospinal fluid and whole blood. Having overcome critical limitations of dielectrophoretic (DEP) operation in physiological media of high conductivity, we utilized target specific DEP techniques to incorporate cell separation, medium exchange, and target concentration into an integrated platform. The proposed microfluidic device can uptake mL volumes of crude biological sample and selectively concentrate target cells into a submicrolitre volume, providing ~10(4) fold of concentration. We designed the device based on the electrokinetic theory and electric field simulation, and tested the device performance with different sample types. The separation efficiency of the device was as high as 97.0% for a bead mixture in TAE buffer and 94.3% and 87.2% for E. coli in human cerebrospinal fluid and blood, respectively. A capture efficiency of 100% was achieved in the concentration chamber. With a relatively simple configuration, the proposed device provides a robust method of continuous sample processing, which can be readily integrated into a fully automated microfluidic diagnostic platform for pathogen detection and quantification.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}

Yi Zhang; Tza-Huei Wang

Quantum Dots-Enabled High-Resolution Analysis of Gene Copy Number Variation Journal Article

IEEE Nanotechnology Magazine, 5 (2), pp. 23-27, 2011, ISSN: 1932-4510.

Links | BibTeX

K. J. Liu; T. D. Rane; Y. Zhang; T. H. Wang

Single-molecule analysis enables free solution hydrodynamic separation using yoctomole levels of DNA Journal Article

J Am Chem Soc, 133 (18), pp. 6898-901, 2011, ISSN: 1520-5126 (Electronic) 0002-7863 (Linking).

Abstract | Links | BibTeX

Y. Zhang; S. Park; K. Liu; J. Tsuan; S. Yang; T. H. Wang

A surface topography assisted droplet manipulation platform for biomarker detection and pathogen identification Journal Article

Lab Chip, 11 (3), pp. 398-406, 2011, ISSN: 1473-0189 (Electronic) 1473-0189 (Linking).

Abstract | Links | BibTeX

2010

Y. Zhang; S. Park; S. Yang; T. H. Wang

An all-in-one microfluidic device for parallel DNA extraction and gene analysis Journal Article

Biomed Microdevices, 12 (6), pp. 1043-9, 2010, ISSN: 1572-8781 (Electronic) 1387-2176 (Linking).

Abstract | Links | BibTeX

V. J. Bailey; B. P. Keeley; C. R. Razavi; E. Griffiths; H. E. Carraway; T. H. Wang

DNA methylation detection using MS-qFRET, a quantum dot-based nanoassay Journal Article

Methods, 52 (3), pp. 237-41, 2010, ISSN: 1095-9130 (Electronic) 1046-2023 (Linking).

Abstract | Links | BibTeX

T. H. Wang; V.J. Bailey; Y. Zhang; K. J. Liu

Quantum Dots DNA Nanosensors - Ultrasensitive Platform for Detecting Genomic Cancer Markers Journal Article

BIOforum Europe, 1-2:28-30, 2010.

BibTeX

X. Jiang; Y. Zheng; H. H. Chen; K. W. Leong; T. H. Wang; H. Q. Mao

Dual-sensitive micellar nanoparticles regulate DNA unpacking and enhance gene-delivery efficiency Journal Article

Adv Mater, 22 (23), pp. 2556-60, 2010, ISSN: 1521-4095 (Electronic) 0935-9648 (Linking).

Links | BibTeX

Tza-Huei Wang; Pak Kin Wong

Transforming Microfluidics into Laboratory Automation Journal Article

Journal of the Association for Laboratory Automation, 15 (3), pp. A15-A16, 2010, ISSN: 15355535.

Links | BibTeX

V. J. Bailey; Y. Zhang; B. P. Keeley; C. Yin; K. L. Pelosky; M. Brock; S. B. Baylin; J. G. Herman; T. H. Wang

Single-tube analysis of DNA methylation with silica superparamagnetic beads Journal Article

Clin Chem, 56 (6), pp. 1022-5, 2010, ISSN: 1530-8561 (Electronic) 0009-9147 (Linking).

Abstract | Links | BibTeX

@article{RN3240,
title = {Single-tube analysis of DNA methylation with silica superparamagnetic beads},
author = { V. J. Bailey and Y. Zhang and B. P. Keeley and C. Yin and K. L. Pelosky and M. Brock and S. B. Baylin and J. G. Herman and T. H. Wang},
url = {https://www.ncbi.nlm.nih.gov/pubmed/20360128},
doi = {10.1373/clinchem.2009.140244},
issn = {1530-8561 (Electronic)
0009-9147 (Linking)},
year = {2010},
date = {2010-06-01},
journal = {Clin Chem},
volume = {56},
number = {6},
pages = {1022-5},
abstract = {BACKGROUND: DNA promoter methylation is a signature for the silencing of tumor suppressor genes. Most widely used methods to detect DNA methylation involve 3 separate, independent processes: DNA extraction, bisulfite conversion, and methylation detection via a PCR method, such as methylation-specific PCR (MSP). This method includes many disconnected steps with associated losses of material, potentially reducing the analytical sensitivity required for analysis of challenging clinical samples. METHODS: Methylation on beads (MOB) is a new technique that integrates DNA extraction, bisulfite conversion, and PCR in a single tube via the use of silica superparamagnetic beads (SSBs) as a common DNA carrier for facilitating cell debris removal and buffer exchange throughout the entire process. In addition, PCR buffer is used to directly elute bisulfite-treated DNA from SSBs for subsequent target amplifications. The diagnostic sensitivity of MOB was evaluated by methylation analysis of the CDKN2A [cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits CDK4); also known as p16(INK4a)] promoter in serum DNA of lung cancer patients and compared with that of conventional methods. RESULTS: Methylation analysis consisting of DNA extraction followed by bisulfite conversion and MSP was successfully carried out within 9 h in a single tube. The median pre-PCR DNA yield was 6.61-fold higher with the MOB technique than with conventional techniques. Furthermore, MOB increased the diagnostic sensitivity in our analysis of the CDKN2A promoter in patient serum by successfully detecting methylation in 74% of cancer patients, vs the 45% detection rate obtained with conventional techniques. CONCLUSIONS: The MOB technique successfully combined 3 processes into a single tube, thereby allowing ease in handling and an increased detection throughput. The increased pre-PCR yield in MOB allowed efficient, diagnostically sensitive methylation detection.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}

K. J. Liu; M. V. Brock; M. Shih Ie; T. H. Wang

Decoding circulating nucleic acids in human serum using microfluidic single molecule spectroscopy Journal Article

J Am Chem Soc, 132 (16), pp. 5793-8, 2010, ISSN: 1520-5126 (Electronic) 0002-7863 (Linking).

Abstract | Links | BibTeX

152 entries « ‹ 3 of 4 › »

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Patents

H. Wang, F.M. Friedrich, J.M. Burke, and K. Liu “Concentration and Accumulation of Target Species in Response to a Gradient of Solute or Solvent” (JHU Ref. C13791; US provisional patent)
H. Wang, K. Hsieh, H. Zec, L. Liu, A.M. Kaushik, Y. Yun, “Multiplexed, Continuous-flow, droplet-based platform for high-throughput genetic detection” (JHU Ref. C13529; US provisional patent)
H. Wang, T.D. Rane, H.C Zec, “System and device for high throughput generation of combinatorial droplets and methods of use” (PCT/US2015/012927; JHU Ref. C12862)
H. Wang, D.J. Shin, “Self-contained cartridge and methods for integrated biochemical assay at the point-of-care” (JHU Ref. C12720)
-H. Wang, S. Yang, M.A. Jacobs, P. Athamanolap, S.I. Fraley, V. Agarwal, V. Parekh. “Melt curve classifier for reliable large-scale genotyping of sequence variants” JHU Case Number C12600, provisional patent
Stephanie I. Fraley, T.-H. Wang, and S. Yang. “A novel technology for broad-based, yet single molecule sensitive profiling in heterogeneous biological samples.” JHU Case Number C12403, provisional patent
H. Wang, Y. Zhang, “Fabrication of Hierarchical Silica Nanomembrane using Heat-Shrinking Polymers and Applications of Silica Nanomembrane for the Solid Phase Extraction of Nucleic Acids”, JHU Case Number C12404, provisional patent
H. Wang, Chi-Hang Chiou, Dong Jin Shin , “Electromagnetically Actuated Droplet Microfluidic Chip and System” (JHU Ref. C12262)
H. Wang and Y. Zhang, “Surface Energy Traps (SETs) Enabled Complex Droplet Manipulation” (JHU Ref. C11971)
H. Wang, K. Liu and Y. Song, “miRNA Analysis Method” (US Provisional US 61/598513, JHU Ref. C11885)
H. Wang, T.D. Rane, H. C. Zec and W.Ch. Chu “Systems and methods for screening a library of samples” (US13/708,510. JHU Ref. C11803)
Yang, T.H. Wang, S.K. Park and Y Zhang, “Method and apparatus for continuous microfluidic sample separation and concentration using AC electric field” (JHU Ref. C11562)
H. Wang, Y. Zhang, S. Park and S. Yang, “Self-sustained Droplet Microfluidic Chip and System for Integrated Sample Preparation and Nucleic Acids Detection” (PCT/US2011/045363, JHU Ref. C11183)
H. Wang, H.Q. Mao, W. Beh and D. Kraitchman, “Kilohertz Generation of Monodisperse Microbeads on a Polymeric Membrane Microfluidic Pseudocheck Valve Device” (PCT/US2011/054598, JHU Ref. C11248)
Ahuja, V. Bailey, S.B. Baylin, J.G. Herman, T.H. Wang, J. Yi, “Early Detection of DNA Methylation Biomarker in Cancer Patient Sera” (PCT/US2011/037926, JHU Ref. C11091)
H. Wang and K. Liu, “Hydrodynamic Particle Separation and Detection Systems and Methods”, (PCT/US2011/056941, JHU Ref. C11263)
H. Wang and K.J. Liu, “Single Molecule Spectroscopy for Analysis of Cell-free Nucleic Acid Biomarkers” (PCT/US2010/033888, JHU Ref. C10750)
H. Wang and Y. Zhang, ” Device and Method of Preparing and Performing Multiple Polymerase Chain Reactions” (US 12/716,031, JHU Ref. C10659)
H. Wang, S. Baylin, J. Herman, H. Easwaran and H. Carraway, “Compositions and Methods for Polynucleotide Extraction and Methylation Detection” (PCT/US2009/000039, JHU Ref. C10249)
M. Ho and T.H. Wang, “Biosensors and Methods for Their Use” (PCT/US2001/025444)
H. Wang, K.J. Liu, and I.M. Shih, ” DNA Integrity Assay (DIA) for Cancer Diagnostics, Using Confocal Fluorescence Spectroscopy” (US8,835,110, JHU Ref. C10534)
H. Wang and K. Liu, “Cylindrical Illumination Confocal Spectroscopy System” (US8,248,609, JHU Ref. C10398)
H. Wang, K.J. Liu, C.M. Puleo and T. Rane, ” Microfluidic System for High-Throughput, Droplet-based, Single Molecule Analysis with Low Reagent Consumption” (US9,284,601, issued on 03/15/2016, JHU Ref. C10662)
F. Miller, J. Huang, T.H. Wang, C.M. Ho and M. Liu, “Electrochemical Detection of Mismatch in Nucleic Acids” (US 7,291,457)
H. Wang, “Method for determining standard cycle time of a stage dynamically” (US5,825,650)
A. Hulbert, J. Herman, M. Brock, T.H. Wang, and A. Stark “Early detection of Individuals at High Risk for Lung Cancer of all Stages Using Gene promoter Methylation in Plasma and Sputum” JHU C13599

Top

Conference Publications

J. Shin, P. Athamanolap, L.Chen, J. Hardick, CA Gaydos and T.H. Wang, “A Smartphone-Based Mobile NAAT Diagnostic Suite for Chlamydia Detection”, Proc. 26^th Anniversary World Congress on Biosensors (Biosensors 2016), (accepted)
M. Friedrich, J.M. Burke, K.J. Liu, T.H. Wang, “In-line Preconcentration, Size Separation, and Single-Molecule Detection Without Applied Electric Fields, Proc. 29th IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS 2016), p. 181-184, 2016
J. Shin, L. Chen and T.H. Wang, “Single-bacteria Confocal Spectroscopy: An Ultrasensitive method for Real-time Monitoring of Bacterial Growth”, Proc. 19th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2015), p. 340-342, 2015
Kaushik, K. Hsieh, L. Chen, D.J. Shin and T.H. Wang, “Rapid Assessment of Bacterial Vitality and Antibiotic Susceptibility via High-Throughput Picoliter-Droplet Single-Cell Assay”, Proc. 19th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2015), p. 531-533, 2015
Liu, K. Hsieh, A. Kaushik, H.C. Zec and T.H. Wang, “Multiplexed, Continuous-Flow, Droplet-Based PCR Genotyping Platform for High-Throughput Agriculture Marker Assisted Selection”, Proc. 19th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2015), p. 1368-1370, 2015
Zec, C. O’Keefe, P. Ma, T.H. Wang, “Ultra-Thin, Evaporation-Resistant PDMS Devices for Absolute Quantification of DNA Using Digital PCR”, The 18th International Conference on Solid-State Sensors, Actuators and Microsystems (Transducers 2015), p.536-539, 2015
Guan, L. Chen, T. Rane, A. Kaushik, T.H. Wang, “Digital Droplet ELOHA For Nucleic Molecule Counting And Analysis”, The 18th International Conference on Solid-State Sensors, Actuators and Microsystems (Transducers 2015), p.536-5460-463, 2015
C. Zec, C.J. Glover, W. Hsieh, L. Liu, C. O’Keefe and T.H. Wang, “Methods for Controlling Water Evaporation in PDMS-Based Microfluidic Devices”, Proc. 18th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2014), p. 1743-1745, 2014
J. Shin, P. Athamanonolap, L. Chen and T.H. Wang, “Integrated droplet microfluidic platform for nucleic acids amplification test of Chlamydia trachomatis infection”, Proc. 24^th Anniversary World Congress on Biosensors (Biosensors 2014)
C. Zec, T.D. Rane, P. Ma and T.H. Wang, “Parallelization of Fission and Fusion- Operations for High Throughput Generation of Combinatorial Droplets”, Proc. 27th IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS 2014), p. 334-337, 2014
J. Shin, L. Chen and T.H. Wang, “A Simple Integrated Diagnostic Platform for DNA Testing of Chlamydia Trachomatis Infection, “ Proc. 17th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2013), p. 1350-1352, 2013
M. Friedrich, K.J. Liu and T.H. Wang, “ Single Molecule Hydrodynamic Separation for Ultrasensitive and Quantitative DNA Size Separations,” Proc. 17th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2013), p. 35-37, 2013
D. Rane, H.C. Zec and T.H. Wang, “A Multiplexed Microfluidic Droplet Platform for Matrix Metalloproteinase Screening”, Proc. 17th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2013), p. 1595-1597, 2013
K. Wu, S. F. Friedrich, K.J. Liu and T.H. Wang, “ Chip-Based DNA Separation in Free Solution by Inertial Hydrodynamic Forces”, Proc. 17th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2013), p. 578-580, 2013
J. Shin, A. Stark and T.H. Wang, “ Droplet Bisulfite Conversion Platform for Epigenetic Cancer Biomarker Detection”, The 17^th International Conference on Solid-State Sensors, Actuators and Microsystems(Transducers 2013), p.2181-2184, 2013
Zhang, Y. Zhang, B. Keeley, A. Stark and T.H. Wang, “Spontaneous Shrinking Silica Nanomembrane for Solid Phase”, Proc. 8th Annual IEEE International Conference on Nano/Micro Engineered and Molecular Systems (IEEE NEMS 2013), p. 444-445, 2013
Athamanolap, B. Keeley, D.J. Shin and T.H. Wang, “Quantitative Analysis of DNA Methylation Based on Melting Curve Analysis”, Proc. 8th Annual IEEE International Conference on Nano/Micro Engineered and Molecular Systems (IEEE NEMS 2013), p.1116-1118, 2013
Zhang, Y Zhang, T.H. Wang, “Hierarchical Silica Nanomembrane Driven by Thermal Shrinkage and its Application for Solid Phase DNA Extraction”, Proc. The 13th IEEE International Conference on Nanotechnology (IEEE NANO 2013), 2013
C. Zec, T.D. Rane, W.C. Chu, V. Wang and T.H. Wang, “ A Microfluidic Droplet Platform for Multiplexed Single Nucleotide Polymorphism Analysis of an Array Plant Genomic DNA Samples”, Proc. 26th IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS 2013), p.263-266, 2013
H. Chiou, D.J. Shin, S. Hosmane, Y Zhang and T.H. Wang, “Electromagnet-Actuated Droplet Platform for Sample-to-Answer Genetic Detection”, Proc. 26th IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS 2013), p.98-101, 2013
Zhang and T.H. Wang, “ All-in-One Droplet Platform for Multiplexed Genetic Detection in Blood” Proc. 26th IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS 2013), p. 1061-1064, 2013
Zhang and T.H. Wang, “Flip-Drop: Droplet Array Created by Surface Energy Trap for Combinatorial Screening,” Proc. 26th IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS 2013), p.1149-1152, 2013
W. Beh, C. Weiss, H.Q. Mao, D. L. Kraitchman, and T.H. Wang, “High-Throughput Microfluidic Preparation of Imaging-Visible Embolic Beads”, Proc. EMBS Micro and Nanotechnology in Medicine Conference, p.59, 2012
Zhang, Y. Zhang, B. Keeley, A. Stark and T.H. Wang, “Fabricating and Applying Hierarchical Silica Nanomembrane with for Solid Phase DNA Extraction” The 6th IEEE International Conference on Nano/Molecular Medicine and Engineering (IEEE NANOMED 2012), 2012
Keeley, Yi Zhang, Ye Zhang, A. Stark, T.H. Wang, “Quantum Dot FRET Linker Probes for Highly Sensitive DNA Methylation Detection”, Proc. IEEE 12^th International Conference on Nanotechnology (IEEE NANO 2012), 7848, p.1-4, 2012
Zhang and T.H. Wang, “Quantum Dot Electrophoretic Mobility Shift Assay and Its Application to the Measurement of Exonuclease Activity” Proc. IEEE 12^th International Conference on Nanotechnology (IEEE NANO 2012), 7685, p.1-4, 2012
Song, L. Zhang, M. Chen and T.H. Wang, “Single Quantum Dot Fluorescence Enhancement by Tunable Nanoporous Gold”, Proc. IEEE 12^th International Conference on Nanotechnology (IEEE NANO 2012), 7877, p.1-4, 2012
Zhang and T.H. Wang, “Droplet Immobilization, Splitting, Metering and Aliquoting with Surface Energy Traps Created Using SU8 Shadow Mask” Proc. 16th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2012), p. 73-75, 2012
Zhang and T.H. Wang, “Surface Energy Trap Assisted Rapid Serial Dilution on Droplet Platform for Bacteria Antibiotics Susceptibility Test” Proc. 16th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2012), p. 467-469, 2012
J. Liu, T.D. Rane, Y. Zhang, C. Beh, D.J. Shin, T.H. Wang, “An Integrated Platform for Single Molecule Free Solution Hydrodynamic Separation Using Yoctomoles of DNA and Picoliter Samples”, ASME 10th International Conference on Nanochannels, Microchannels and Minichannels (ICNMM 2012), ICNMM2012-73154 (p.1-6), 2012
Zec, T.D. Rane, W.C. Chu and T.H. Wang, “Multiplexed Screening of a Large Library of Biological Samples through on-Demand Droplet Generation and Fusion”, ASME 2012 10th International Conference on Nanochannels, Microchannels and Minichannels (ICNMM 2012), ICNMM2012-73159 (p.1-6), 2012
Song, Y. Zhang, and T.H. Wang, “Single Quantum Dot-Based Multiplexed Point Mutation Detection by Gap Ligase Chain Reaction”, Proc. 15th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2011), p.1779-1781, 2011
Zhang, D.J. Shin and T.H. Wang, “Detecting Genetic Variations in A Droplet”, Proc. 15th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2011), p.1179-1181, 2011
J. Liu and T.H. Wang, “ PCR-free, microfluidic single molecule analysis of circulating nucleic acids in lung cancer patient serum”, Proc. 33rd Annual International Conference of the IEEE Engineering in Medicine and Biology Society (EMBC ’11), p.8392-8395, 2011
Zhang and T.H. Wang, “ An Active Gyroscopic Magnetic Micromixer for Rapid Fluid Mixing in Droplet Based Microfluidic Systems”, The 16^th International Conference on Solid-State Sensors, Actuators and Microsystems(Transducers 2011), p.1769-1772, 2011
Zhang, S.K. Park, S. Yang and T.H. Wang, “Fully Integrated Droplet Based Point-of-Care Platform for Molecular Detection from Crude Biosamples”, The 16^th International Conference on Solid-State Sensors, Actuators and Microsystems(Transducers 2011), p.1927-1930, 2011
H. Wang, V. Bailey and K. Liu, “Quantum Dots and Microfluidic Single Molecule Detection for Screening Genetic and Epigenetic Cancer Markers in Clinical Samples”, Proc. 2011 SPIE Defense, Security and Sensing Conference, Volume 8031, P. 80311W
D. Rane, H. Zec, C.M. Puleo, A.P. Lee and T.H. Wang, “High-Throughput Single-Cell Pathogen Detection on a Droplet Microfluidic Platform”, Proc. 24th IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS 2011), p.881-884, 2011
Stark, Y. Zhang, V. Bailey, B. Keeley. T.H. Wang, “Increasing Throughput and Sensitivity of DNA Methylation Analysis through Functional Nanoparticles” The 6th Annual IEEE International Conference on Nano/Micro Engineered and Molecular Systems (IEEE NEMS 2011), 1091-1094, 2011
Y Zhang and T.H. Wang, “A Quantum Dot Based Electrophoretic Mobility Shift Assay for High Resolution Copy Number Variation Study”, The 6th Annual IEEE International Conference on Nano/Micro Engineered and Molecular Systems (IEEE NEMS 2011), p.841-844, 2011
Zhang, I.M. Shih, T.L. Wang and T.H. Wang, ” A Quantum Dot Based Nanoassay for Quantifying Gene Copy Number with Ultrahigh Resolution”, Proc. 14th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2010), p.1154-1156, 2010
W. Beh, W. Zhou and T.H. Wang, “Oxygen Plasma-Free Microfluidic Device Sealing”, Proc. 14th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2010) p.1217-1219, 2010
W. Beh, D. Kraitchman, H.Q. Mao, T.H. Wang, “High-throughput Monodisperse Alginate Gel Bead Formation using Microfluidic Pseudo-Check Valve”, Proc. 14th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2010) p.425-427, 2010
Zhang and T.H. Wang, “An Automated All-in-one Microfluidic Device for Parallel Solid Phase DNA Extraction and Droplet-in-Oil PCR Analysis”, Proc. 23rd IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS 2010), P.971-974, 2010
Zhang and T.H. Wang, “Geomorphology-Assisted Manopulation of Magnet-Actuated Droplet for Solid Phase DNA Extraction and Droplet-in-Oil PCR”, Proc. 23rd IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS 2010), p.1047-1050, 2010
D. Rane, C.M. Puleo, H. Zec, Y. Zhang, A.P. Lee and T.H. Wamh, “Analyte Detection in Droplets: One Molecule at a Time”, Proc. 13th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2009), p.15-17, 2009
M. Puleo, H.C. Zec, Y. Sung and T.H. Wang, “Micro-evaporator as Interconnects to Low-Volume Microfluidic Components”, Proc. 13th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2009), p.932-935, 2009
J. Bailey, C.M. Puleo, Y.P. Ho, H.C. Yeh, T.H. Wang, “Quantum Dots in Molecular Detection of Diseases”, 31st Annual International Conference on the IEEE Engineering in Medicine and Biology Society ( IEEE EMBC 2009), p. 4089-4092, 2009
Zhang, V. Bailey, C.M. Puleo, H. Easwaran, E. Griffiths, J.G. Herman, S.B. Baylin, T.H. Wang, “High Throughput DNA Methylation Analysis on a Droplet-in-Oil Polymerase Chain Reaction Array”, The 15^th International Conference on Solid-State Sensors, Actuators and Microsystems(Transducers 2009), p.806-808, 2009
Zhang, V. Bailey, C.M. Puleo, C. Chen and T.H. Wang, “Multiple gene Analysis within a Simple Droplet-in-Oil Microfluidic PCR Platform”, Proc. 12th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2008), p. 751-753 , 2008
P. Ho, H.H. Chen, K. Leong and T.H. Wang “Quantitative Kinetic Analysis of DNA Nanocomplex Self-Assembly with Quantum Dots FRET in a Microfluidic Device”, Proc. 21st IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS 2008), p. 30-33, 2008
M. Puleo, H.C. Yeh and T.H. Wang “Single Molecule Detection in Truly, Nanoliter-Sized Volumes: Coupling Evaporation-Based, Microfluidic Concentration with Confocal Fluorescence Spectroscopy” Proc. 21st IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS 2008), p.200-203, 2008
Liu and T.H. Wang, “Quantitative Confocal Spectroscopy – Rectifying the Limitations of Single Molecule Detection”, Proc. 3^rd Annual IEEE International Conferences on Nano/Micro Engineered and Molecular Systems (IEEE-NEMS 2008), p. 1189-1192, 2008
C. Yeh. Y.P. Ho, C.M. Puleo and T.H. Wang,” Towards single-molecule diagnostics using microfluidic manipulation and quantum dot nanosensors, Proc. 5^th International Conference on Nanochannels, Microchannels and Minichannels, (ICNMM 2007), p. 1133-1140, 2007
P. Ho and T.H. Wang, “Multiplexed Detection of Anthrax Sequences with Quantum Dot Nanoprobes”, Proc. IEEE/NLM Life Science Systems and Application Workshop, p. 62-63, 2006
Murray , K. Rebello, J. Crookston, J. Miragliotta and T.H. Wang, “High-degree Concentration of Bio-agents Using Electrokinetic Manipulations”, Proc. IEEE/NLM Life Science Systems and Application Workshop, p. 94-95, 2006
P. Ho, M.C. Kung and T.H. Wang, ” Separation-free Detection of Low-abundant Biomolecules with Two-Color Colocalization of Quantum Dot Probes,” Proc. 9th International Conference on Miniaturized Chemical and Biochemical Analysis Systems (micro-TAS 2005), p. 1330-1332, 2005
C. Yeh, Y.P. Ho and T.H. Wang, “Quantum Dot-Mediated Separation-Free Assay for Point Mutation Detection ” Proc. NSTI- Bio-Nanotechnology Conference, 198-201, 2005
C. Yeh, E. Simone, C.Y. Zhang and T.H. Wang, “Single Bio-Molecule Detection with Quantum-dots in Flow-rate Controlled Microchannel”, Proc. 17h IEEE Annual Workshop of Micro Electro Mechanical Systems (IEEE MEMS 2004), p 371-374, 2004
Y. Chao, C.Y. Zhang and T.H. Wang,” Measurement of in-situ Flow Velocity Using Single-Molecule Detection for the Application of Biomolecule Quantification”, Proc. Hilton Head 2004 Solid-State Sensor, Actuator, and Microsystems Workshop (Hilton Head 2004), p. 176-179, 2004
H. Wang and C.M. Ho, “Nano/micro Te
chnologies for Single Molecule Manipulation and Detection”, IEEE International Conference on Robotics and Control (IEEE ICRA 2003), vol. 3, p. 3630-3635, 2003
K. Wong, C.Y. Chen, T.H. Wang and C.M. Ho, “An AC Electroosmotic Processor for Biomolecules,” Proc. 12^th International Conference on Solid-state Sensors, Actuators, and Microsystems (Transducers 2003), vol. 1, p. 20-23, 2003
K. Wong, T.H. Wang and C.M. Ho, “Optical Fiber Tip Fabricated by Surface Tension Controlled Etching”, Proc. Hilton Head 2002: Solid-State Sensor, Actuator, and Microsystems Workshop (Hilton Head 2002), p. 94-97, 2002
H. Wang, P.K. Wong and C.M. Ho, “Electrical Molecular Focusing for Laser Induced Fluorescence Based Single DNA Detection”, Proc. 15^th IEEE International Conference on Micro Electro Mechanical Systems(IEEE MEMS 2002), p. 15-18, 2002
H. Wang, S. Masset and C.M. Ho, “A Zepto Mole DNA Micro Sensor,” Proc. 14^th IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS), p. 431-434, 2001
H. Wang, S. Masset and C.M. Ho “Molecular Beacon Based Biological Detection System”, Proc. The International Conference on Mathematics and Engineering Techniques in Medicine and Biological Science, p. 295-300, 2000
H. Wang, K.C. Lin and S.R. Huang “Method of Dynamically Determining Cycle Time of a Working Stage”, Proc. 21^st IEEE/CPMT International Electronics manufacturing Technology Symposium, p. 403-407, 1997
C. Chang, W.L. Jan. T.H. Wang and C.S. Chang, “Analysis of Proportional Machine Allocation in a Deterministic Re-entrant Line”, Proc. INRIA/IEEE Conference on Emerging Technology and Factory Automation, vol. 2, p. 1-5, 1995