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.