Establishment of fluorescence RT-RAA detection method for influenza A virus
Objective To establish a rapid detection method for influenza A virus using reverse transcription recombi-nase-aided isothermal amplification(RT-RAA).Methods Firstly,the gene sequences of influenza A viruses were searched on NCBI and compared using DNAstar software.Three pairs of primers and one fluorescent probe were de-signed targeted conserved M gene to establish fluorescent RT-RAA assay for influenza A virus.The established RT-RAA assay was also tested for specificity,sensitivity and reproducibility as well as for clinical samples.Results One set of primer and probe with good amplification effect had been successfully screened,and fluorescent RT-RAA method for the rapid detection of influenza A virus was established,which could complete the detection by constant amplification at 42℃ for 20 min.A minimum of 3.18×100 copies/μl of influenza A virus recombinant plasmid was detected and there was no cross reaction with influenza B virus,SARS-CoV-2 and dengue virus.Twenty influenza A virus nucleic acid positive clinical samples were tested using commercial kit and the RT-RAA method established in this study,the compliance rate of the two methods was 100％.Conclusion A fluorescent RT-RAA method was suc-cessfully established for the rapid detection of influenza A virus with good specificity,sensitivity,providing a new method for influenza A virus detection.
Influenza A virusRecombinase-aided isothermal amplificationDetection
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