Development of a multi‐recombinase polymerase amplification assay for rapid identification of COVID‐19, influenza A and B

作者：L. Liang, Miaojin Zhu, Rui He, Danrong Shi, Rui Luo, Jia Ji, Linfang Cheng, Xiang‐Yun Lu, Wei Lü, Fu‐Ming Liu, Zhigang Wu, Nanping Wu, Hang Chen, Zhe Chen, Hangping Yao · 发表于：Journal of Medical Virology · 年份：2022 · DOI：10.1002/jmv.28139 · 被引用次数：22 · 研究领域：SARS-CoV-2 detection and testing、Respiratory viral infections research、SARS-CoV-2 and COVID-19 Research

Abstract The coronavirus disease 2019 (COVID‐19) pandemic caused extensive loss of life worldwide. Further, the COVID‐19 and influenza mix‐infection had caused great distress to the diagnosis of the disease. To control illness progression and limit viral spread within the population, a real‐time reverse‐transcription PCR (RT‐PCR) assay for early diagnosis of COVID‐19 was developed, but detection was time‐consuming (4–6 h). To improve the diagnosis of COVID‐19 and influenza, we herein developed a recombinase polymerase amplification (RPA) method for simple and rapid amplification of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS‐CoV‐2), the causative agent of COVID‐19 and Influenza A (H1N1, H3N2) and B (influenza B). Genes encoding the matrix protein (M) for H1N1, and the hemagglutinin (HA) for H3N2, and the polymerase A (PA) for Influenza B, and the nucleocapsid protein (N), the RNA‐dependent‐RNA polymerase (RdRP) in the open reading frame 1ab (ORF1ab) region, and the envelope protein (E) for SARS‐CoV‐2 were selected, and specific primers were designed. We validated our method using SARS‐CoV‐2, H1N1, H3N2 and influenza B plasmid standards and RNA samples extracted from COVID‐19 and Influenza A/B (RT‐PCR‐verified) positive patients. The method could detect SARS‐CoV‐2 plasmid standard DNA quantitatively between 10 2 and 10 5 copies/ml with a log linearity of 0.99 in 22 min. And this method also be very effective in simultaneous detection of H1N1, H3N2 and influenza B. Cl...