SD-chip enabled quantitative detection of HIV RNA using digital nucleic acid sequence-based amplification (dNASBA)

被引：44

作者：

Wang, Jiasi ^{[1
]}

Kreutz, Jason E. ^{[1
]}

Thompson, Alison M. ^{[2
]}

Qin, Yuling ^{[1
]}

Sheen, Allison M. ^{[1
]}

Wang, Jingang ^{[1
]}

Wu, Li ^{[1
]}

Xu, Shihan ^{[1
]}

Chang, Ming ^{[5
]}

Raugi, Dana N. ^{[3
]}

Smith, Robert A. ^{[3
]}

Gottlieb, Geoffrey S. ^{[3
,4
]}

Chiu, Daniel T. ^{[1
]}

机构：

[1] Univ Washington, Dept Chem, Seattle, WA 98195 USA

[2] Fred Hutchinson Canc Res Ctr, Seattle, WA 98109 USA

[3] Univ Washington, Dept Med, Ctr Emerging & Reemerging Infect Dis, Div Allergy & Infect Dis, Seattle, WA USA

[4] Univ Washington, Dept Global Hlth, Seattle, WA 98195 USA

[5] Univ Washington, Dept Lab Med, Seattle, WA 98195 USA

来源：

LAB ON A CHIP | 2018年 / 18卷 / 22期

基金：

美国国家卫生研究院; 比尔及梅琳达.盖茨基金会;

关键词：

VIRUS TYPE-1 RNA; MESSENGER-RNA; SELF-DIGITIZATION; VIRAL LOAD; NASBA; QUANTIFICATION; PLASMA; INFECTION; TECHNOLOGY; DIAGNOSIS;

D O I：

10.1039/c8lc00956b

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：3501 / 3506

页数：6

共 39 条

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