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Development of an Internally Controlled Reverse Transcription Recombinase-aided Amplification Assay for the Rapid and
Guo Hao Fan1, Xin Xin Shen2, Fan Li3
1Department of Core Facility, Ministry of Health Key Laboratory of Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China; Biosafety 3 Laboratory, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Abstract:
West Nile virus (WNV) causes West Nile fever and West Nile encephalitis. Because infection by WNV creates serious public health problems, its simple, rapid, and visual detection is very important in clinical practice, especially in resource-limited laboratories. We have developed a rapid, specific, and highly sensitive internally controlled reverse transcription recombinase-aided amplification (RTRAA) assay to detect WNV, using both real-time fluorescence and the lateral flow dipstick (LFD) at 39.0 °C for 30 min. The analytical sensitivity of the RT-RAA assay was 10 plasmid copies and 1.6 pfu per reaction with real-time fluorescence, and 1,000 plasmid copies per reaction with the LFD. No crossreaction with other control viruses was observed. Compared with the RT-qPCR assay, the RT-RAA assay demonstrated 100% sensitivity and 100% specificity for WNV.
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