دورية أكاديمية
A microfluidic-based quantitative analysis system for the multiplexed genetic diagnosis of human viral infections using colorimetric loop-mediated isothermal amplification.
| العنوان: | A microfluidic-based quantitative analysis system for the multiplexed genetic diagnosis of human viral infections using colorimetric loop-mediated isothermal amplification. |
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| المؤلفون: | Natsuhara D; Department of Mechanical Engineering, Toyohashi University of Technology, Aichi 441-8580, Japan. d-natsuhara@mems.me.tut.ac.jp., Miyajima A; Department of Mechanical Engineering, Toyohashi University of Technology, Aichi 441-8580, Japan. d-natsuhara@mems.me.tut.ac.jp., Bussho T; Department of Mechanical Engineering, Toyohashi University of Technology, Aichi 441-8580, Japan. d-natsuhara@mems.me.tut.ac.jp., Okamoto S; Department of Mechanical Engineering, Toyohashi University of Technology, Aichi 441-8580, Japan. d-natsuhara@mems.me.tut.ac.jp., Nagai M; Department of Mechanical Engineering, Toyohashi University of Technology, Aichi 441-8580, Japan. d-natsuhara@mems.me.tut.ac.jp.; Institute for Research on Next-generation Semiconductor and Sensing Science (IRES2), Toyohashi University of Technology, Aichi 441-8580, Japan., Ihira M; Faculty of Clinical Science for Biological Monitoring, Fujita Health University, Aichi 470-1192, Japan., Shibata T; Department of Mechanical Engineering, Toyohashi University of Technology, Aichi 441-8580, Japan. d-natsuhara@mems.me.tut.ac.jp. |
| المصدر: | The Analyst [Analyst] 2024 Jun 10; Vol. 149 (12), pp. 3335-3345. Date of Electronic Publication: 2024 Jun 10. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Royal Society of Chemistry Country of Publication: England NLM ID: 0372652 Publication Model: Electronic Cited Medium: Internet ISSN: 1364-5528 (Electronic) Linking ISSN: 00032654 NLM ISO Abbreviation: Analyst Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Cambridge : Royal Society of Chemistry Original Publication: London : Chemical Society |
| مواضيع طبية MeSH: | Nucleic Acid Amplification Techniques*/methods , Nucleic Acid Amplification Techniques*/instrumentation , Colorimetry*/methods , Colorimetry*/instrumentation , Molecular Diagnostic Techniques*/methods , Molecular Diagnostic Techniques*/instrumentation , Lab-On-A-Chip Devices*, Humans ; Microfluidic Analytical Techniques/instrumentation ; Microfluidic Analytical Techniques/methods ; DNA, Viral/analysis ; DNA, Viral/genetics ; Virus Diseases/diagnosis ; Limit of Detection |
| مستخلص: | In this study, a microfluidic-based system utilizing colorimetric loop-mediated isothermal amplification (LAMP) is introduced for the quantitative analysis of nucleic acid targets. This system offers a user-friendly and cost-effective platform for the multiplexed genetic diagnosis of various infectious diseases across multiple samples. It includes time-lapse imaging equipment for capturing images of the microfluidic device during the LAMP assay and a hue-based quantitative analysis software to analyze the LAMP reaction, streamlining diagnostic procedures. An electric pipette was used to simplify the loading of samples and LAMP reagents into the device, allowing easy operation even by untrained individuals. The hue-based analysis software employs efficient image processing and post-processing techniques to calculate DNA amplification curves based on color changes in multiple reaction chambers. This software automates several tasks, such as identifying reaction chamber areas from time-lapse images, quantifying color information within each chamber, correcting baselines of DNA amplification curves, fitting experimental data to theoretical curves, and determining the threshold time for each curve. To validate the developed system, conventional off-chip LAMP assays were conducted with a 25 μL reaction mixture in 0.2 mL polymerase chain reaction (PCR) tubes using a real-time turbidimeter. The results indicated that the threshold time obtained using the colorimetric LAMP assay in the developed system is comparable to that obtained with real-time turbidity measurements in PCR tubes, demonstrating the system's capability for quantitative analysis of target nucleic acids, including those from human herpesviruses. |
| المشرفين على المادة: | 0 (DNA, Viral) |
| SCR Protocol: | LAMP assay |
| تواريخ الأحداث: | Date Created: 20240502 Date Completed: 20240610 Latest Revision: 20240610 |
| رمز التحديث: | 20240610 |
| DOI: | 10.1039/d4an00215f |
| PMID: | 38695841 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1364-5528 |
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| DOI: | 10.1039/d4an00215f |