Digital droplet RT-LAMP increases speed of SARS-CoV-2 viral RNA detection

被引：2

作者：

Yuan, Yuan ^{[1
,2
]}

Ellis, Perry ^{[2
]}

Tao, Ye ^{[2
]}

Bikos, Dimitri A. ^{[3
,4
]}

Loveday, Emma K. ^{[3
,4
]}

Thomas, Mallory M. ^{[3
,4
]}

Wilking, James N. ^{[3
,4
,5
]}

Chang, Connie B. ^{[3
,4
,5
]}

Ye, Fangfu ^{[1
,6
,11
]}

Weitz, David A. ^{[2
,7
,8
,9
,10
]}

机构：

[1] Univ Chinese Acad Sci, Wenzhou Inst, Oujiang Lab, Zhejiang Lab Regenerat Med Vis & Brain Hlth, Wenzhou, Zhejiang, Peoples R China

[2] Harvard Univ, John A Paulson Sch Engn & Appl Sci, Cambridge, MA 02138 USA

[3] Montana State Univ, Dept Chem & Biol Engn, Bozeman, MT USA

[4] Montana State Univ, Ctr Biofilm Engn, Bozeman, MT USA

[5] Mayo Clin, Dept Physiol & Biomed Engn, Rochester, MN USA

[6] Chinese Acad Sci, Beijing Natl Lab Condensed Matter Phys, Inst Phys, Beijing, Peoples R China

[7] Harvard Univ, Dept Phys, Cambridge, MA 02138 USA

[8] Harvard Univ, Wyss Inst Biol Inspired Engn, Boston, MA USA

[9] Harvard Univ, Dept Phys, Phys & Appl Phys, Pierce Hall 231,29 Oxford St, Cambridge, MA 02138 USA

[10] Harvard Univ, SEAS, Pierce Hall 231,29 Oxford St, Cambridge, MA 02138 USA

[11] Chinese Acad Sci, Inst Phys, 8 Zhongguancun 3rd South St, Beijing 100190, Peoples R China

来源：

SMART MEDICINE | 2024年 / 3卷 / 02期

基金：

美国国家卫生研究院; 中国国家自然科学基金;

关键词：

ddRT-LAMP; droplet microfluidics; droplet size; RNA rapid detection; SARS-CoV-2; virus; ACCURACY; PCR;

D O I：

10.1002/SMMD.20240008

中图分类号：

R318 [生物医学工程];

学科分类号：

0831 ;

摘要：

Nucleic acid amplification testing (NAAT) remains one of the most reliable methods for pathogen identification. However, conventional bulk NAATs may not be sufficiently fast or sensitive enough for the detection of clinically-relevant pathogens in point-of-care testing. Here, we have developed a digital droplet RT-LAMP (ddRT-LAMP) assay that rapidly and quantitatively detects the SARS-CoV-2 viral E gene in microfluidic drops. Droplet partitioning using ddRT-LAMP significantly accelerates detection times across a wide range of template concentrations compared to bulk RT-LAMP assays. We discover that a reduction in droplet diameter decreases assay times up to a certain size, upon which surface adsorption of the RT-LAMP polymerase reduces reaction efficiency. Optimization of drop size and polymerase concentration enables rapid, sensitive, and quantitative detection of the SARS-CoV-2 E gene in only 8 min. These results highlight the potential of ddRT-LAMP assays as an excellent platform for quantitative point-of-care testing. We have developed a digital droplet RT-LAMP (ddRT-LAMP) assay that rapidly and quantitatively detects the SARS-CoV-2 viral E gene in microfluidic drops. Droplet partitioning using ddRT-LAMP significantly accelerates detection times across a wide range of template concentrations compared to bulk RT-LAMP assays. image

引用

页数：9

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