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bioRxiv · 10.1101/2020.10.22.350025

Reverse-transcription recombinase-aided amplification assay for H5 subtype avian influenza virus

Wang, K.·Wang, S.·Li, Y.·Jiang, N.·Zhang, F.·Zhuang, Q.·Hou, G.·Jiang, L.·Yu, J.·Yu, X.·Liu, H.·Zhao, C.·Yuan, L.·Huang, B.
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Abstract

The H5 subtype Avian Influenza Virus has caused huge economic losses to the poultry industry and is a threat to human health. A rapid and simple test is needed to confirm infection in suspected cases during disease outbreaks. In this study, we developed a reverse-transcription recombinase-aided amplification assay for the detection of H5 subtype avian influenza virus. Assays were performed at a single temperature (39{degrees}C), and the results were obtained within 20 min. The assay showed no cross-detection with Newcastle disease virus or infectious bronchitis virus. The analytical sensitivity was 103 RNA copies per reaction at a 95% confidence interval according to probit regression analysis, with 100% specificity. Compared with published reverse-transcription quantitative real-time polymerase chain reaction assays, the {kappa} value of the reverse transcription recombinase-aided amplification assay in 365 avian clinical samples was 0.970 (p < 0.001). The sensitivity for avian clinical sample detection was 94.44% (95%CI, 70.63% - 99.71%), and the specificity was 100% (95%CI, 98.64% - 100%). These results indicated that our reverse-transcription recombinase-aided amplification assay may be a valuable tool for detecting H5 subtype avian influenza virus.

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Wang, K., Wang, S., Li, Y., Jiang, N., Zhang, F., Zhuang, Q., Hou, G., Jiang, L., Yu, J., Yu, X., Liu, H., Zhao, C., Yuan, L., Huang, B.. 2020-10-22. Reverse-transcription recombinase-aided amplification assay for H5 subtype avian influenza virus. https://doi.org/10.1101/2020.10.22.350025

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