SD-chip enabled quantitative detection of HIV RNA using digital nucleic acid sequence-based amplification (dNASBA)
| Autor: | Robert A. Smith, Li Wu, Jason E. Kreutz, Geoffrey S. Gottlieb, Jingang Wang, Ming Chang, Dana N. Raugi, Jiasi Wang, Yuling Qin, Alison M. Thompson, Daniel T. Chiu, Allison M. Sheen, Shihan Xu |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Computer science Nucleic acid sequence based amplification Biomedical Engineering Bioengineering Computational biology 010402 general chemistry 01 natural sciences Biochemistry Article 03 medical and health sciences Viral genetics Lab-On-A-Chip Devices parasitic diseases Humans Self-Sustained Sequence Replication Plasma samples Nucleic acid sequence RNA General Chemistry Chip NASBA 0104 chemical sciences 030104 developmental biology HIV-1 RNA Viral Viral load |
| Zdroj: | Lab on a Chip. 18:3501-3506 |
| ISSN: | 1473-0189 1473-0197 |
| Popis: | Quantitative detection of RNA is important in molecular biology and clinical diagnostics. Nucleic acid sequence-based amplification (NASBA), a single-step method to amplify single-stranded RNA, is attractive for use in point-of-care (POC) diagnostics because it is an isothermal technique that is as sensitive as RT-PCR with a shorter reaction time. However, NASBA is limited in its ability to provide accurate quantitative information, such as viral load or RNA copy number. Here we test a digital format of NASBA (dNASBA) using a self-digitization (SD) chip platform, and apply it to quantifying HIV-1 RNA. We demonstrate that dNASBA is more sensitive and accurate than the real-time quantitative NASBA, and can be used to quantify HIV-1 RNA in plasma samples. Digital NASBA is thus a promising POC diagnostics tool for use in resource-limited settings. |
| Databáze: | OpenAIRE |
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