Frontiers in Microbiology (Feb 2021)

One-Step Reverse-Transcription Recombinase Polymerase Amplification Using Lateral Flow Strips for the Detection of Coxsackievirus A6

  • Jia Xie,
  • Xiaohan Yang,
  • Xiaohan Yang,
  • Lei Duan,
  • Keyi Chen,
  • Keyi Chen,
  • Pan Liu,
  • Pan Liu,
  • Wenli Zhan,
  • Wenli Zhan,
  • Changbin Zhang,
  • Changbin Zhang,
  • Hongyu Zhao,
  • Hongyu Zhao,
  • Mengru Wei,
  • Yuan Tang,
  • Mingyong Luo,
  • Mingyong Luo

DOI
https://doi.org/10.3389/fmicb.2021.629533
Journal volume & issue
Vol. 12

Abstract

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Hand, foot, and mouth disease (HFMD) is a common infectious disease affecting mainly children under 5 years of age. Coxsackievirus A6 (CVA-6), a major causative pathogen of HFMD, has caused outbreaks in recent years. Currently, no effective vaccine or antiviral treatments are available. In this study, one-step reverse-transcription recombinase polymerase amplification (RT-RPA), combined with a disposable lateral flow strip (LFS) assay, was developed to detect CVA-6. This assay can be performed in less than 35 min at 37°C without expensive instruments, and the result can be observed directly with the naked eye. The sensitivity of the RT-RPA-LFS was 10 copies per reaction, which was comparable to that of the conventional real-time quantitative polymerase chain reaction (qPCR) assays. Moreover, the assay specificity was 100%. The clinical performance of the RT-RPA-LFS assay was evaluated using 142 clinical samples, and the coincidence rate between RT-RPA-LFS and qPCR was 100%. Therefore, our RT-RPA-LFS assay provides a simple and rapid approach for point-of-care CVA-6 diagnosis.

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