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Fluorescent reverse transcription recombinase-aided amplification assay for sensitive and rapid detection of six respiratory viruses
  • +6
  • Xudong Guo,
  • Dongli Gao,
  • Minghui Song,
  • Yi Yang,
  • Dan Sun,
  • Wanying Liu,
  • Hua Shi,
  • Rongtao Zhao,
  • Hongbin Song
Xudong Guo
Chinese PLA Center for Disease Control and Prevention
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Dongli Gao
Chinese PLA Center for Disease Control and Prevention
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Minghui Song
Hainan Hospital of Chinese PLA General Hospital
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Yi Yang
Chinese PLA Center for Disease Control and Prevention
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Dan Sun
Hainan Hospital of Chinese PLA General Hospital
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Wanying Liu
Chinese PLA Center for Disease Control and Prevention
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Hua Shi
Chinese PLA Center for Disease Control and Prevention
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Rongtao Zhao
Chinese PLA Center for Disease Control and Prevention

Corresponding Author:[email protected]

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Hongbin Song
Chinese PLA Center for Disease Control and Prevention
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Abstract

Background: Rapid detection of pathogens is important for the timely control of outbreaks, especially for respiratory infectious diseases that are prone to spread and outbreaks. Methods: In this work, we developed a sensitive reverse transcription recombinase-aided amplification (RT-RAA) assay for the rapid detection of six common respiratory viruses including respiratory syncytial virus type A (RSVA), influenza A virus (Flu A), influenza B virus (Flu B), human parainfluenza virus (HPIV), SARS-CoV-2 and adenovirus (ADV). The nucleic acid standards and pharyngeal swab samples were used to test the sensitivity, specificity, reliability of the established RAA assay. Results: The assay could be completed within 20 minutes at 39℃ using a portable built-in power device. The detection limits for the six viruses were all less than 1000 copies/mL and reached 10 copies/mL for ADV. Excellent specificity was demonstrated by cross-testing with 21 different pathogen nucleic acids. The results of RT-RAA and RT-PCR were consistent in 85 laboratory-conserved pharyngeal swab samples, but RT-RAA was more time-saving and portable. Meanwhile, the RT-RAA assay using the same test procedure for six viruses could allow operators the flexibility to select the number of samples and pathogens to be detected in one test. Conclusions: This portable, sensitive and reliable RT-RAA assay for rapid detection of multiple respiratory viruses could be applied to health resource-poor areas and outbreak sites.