Fluorescentreverse transcription recombinase-aided amplification assay for sensitive andrapid detection of six respiratory viruses
Xudong Guo1†, Dongli Gao1,2, Minghui Song3†, Yi Yang1, Dan Sun3, Wanying Liu1, Hua Shi1, Rongtao Zhao1,*, Hongbin Song1,2,*
1Chinese PLA Center for Disease Control and Prevention, Beijing 100071, China
2College of Public Health, Zhengzhou University, Henan 450001, China
3 Hainan Hospital of Chinese PLA General Hospital, Hainan 572013, China
These authors contributed equally to this work
*Corresponding author:
Rongtao Zhao, Email: zhaorongtao1984@163.com;
Hongbin Song, Email: hongbinsong@263.net
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 103 copies/mL and reached 101copies/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.