Exploratory research on detection of influenza viruses by reverse transcriptase-recombinase polymerase amplification

Abstract

Objective To establish and hevaluate a detection method for influenza virus using reverse transcriptase-recombinase polymerase amplification (RT-RPA). Methods RT-RPA was developed for detection of influenza viruses (type A and B) and subtyping of H1 and H3 using the primers targeted matrix and hemagglutinin (HA) genes of influenza A virus, and non-structural (NS) protein gene of influenza B virus. The specificity and sensitivity of RT-RPA were determined. Results The RT-RPA for the detection of influenza viruses showed specific amplification products of corresponding target gene, but no amplification products for other respiratory viruses, indicating that the method had good specificity. The detection limits of RT-RPA were 100 copies/μl. RT-RPA combined with SYBR Green I was used for the detection of influenza B virus with the detection limit of 100 copies/μl. Conclusions The feasibility of detecting influenza virus by RT-RPA was preliminarily confirmed. Key words: Influenza virus; Detection; Subtyping; Recombinase polymerase amplification