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Microfluidic Chip with Two-Stage Isothermal Amplification Method for Highly Sensitive Parallel Detection of SARS-CoV-2 and Measles Virus.
Huang, Qin; Shan, Xiaohui; Cao, Ranran; Jin, Xiangyu; Lin, Xue; He, Qiurong; Zhu, Yulei; Fu, Rongxin; Du, Wenli; Lv, Wenqi; Xia, Ying; Huang, Guoliang.
  • Huang Q; Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
  • Shan X; Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
  • Cao R; Sichuan Center for Disease Control and Prevention, Chengdu 610041, China.
  • Jin X; Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
  • Lin X; Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
  • He Q; West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu 610041, China.
  • Zhu Y; West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu 610041, China.
  • Fu R; Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
  • Du W; Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
  • Lv W; Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
  • Xia Y; West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu 610041, China.
  • Huang G; Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China.
Micromachines (Basel) ; 12(12)2021 Dec 19.
Article in English | MEDLINE | ID: covidwho-1580576
ABSTRACT
A two-stage isothermal amplification method, which consists of a first-stage basic recombinase polymerase amplification (RPA) and a second-stage fluorescence loop-mediated isothermal amplification (LAMP), as well as a microfluidic-chip-based portable system, were developed in this study; these enabled parallel detection of multiplex targets in real time in around one hour, with high sensitivity and specificity, without cross-contamination. The consumption of the sample and the reagent was 2.1 µL and 10.6 µL per reaction for RPA and LAMP, respectively. The lowest detection limit (LOD) was about 10 copies. The clinical amplification of about 40 nasopharyngeal swab samples, containing 17 SARS-CoV-2 (severe acute respiratory syndrome coronavirus) and 23 measles viruses (MV), were parallel tested by using the microfluidic chip. Both clinical specificity and sensitivity were 100% for MV, and the clinical specificity and sensitivity were 94.12% and 95.83% for SARS-CoV-2, respectively. This two-stage isothermal amplification method based on the microfluidic chip format offers a convenient, clinically parallel molecular diagnostic method, which can identify different nucleic acid samples simultaneously and in a timely manner, and with a low cost of the reaction reagent. It is especially suitable for resource-limited areas and point-of-care testing (POCT).
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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Randomized controlled trials Language: English Year: 2021 Document Type: Article Affiliation country: Mi12121582

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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Randomized controlled trials Language: English Year: 2021 Document Type: Article Affiliation country: Mi12121582