Four multiplex RT-PCR for rapid detection of 2013 novel avian influenza A(H7N9) virus

Objective To establish a four multiplex RT-PCR assay for rapid and accurate detection of 2013 novel avian influenza A(H7N9) virus in throat-swab specimens of patients with fever.Methods The universal primers for amplifying the M gene were designed for detection of influenza A viruses. Two pairs of specific primers of HA and NA gene were designed to detect the 2013 novel A(H7N9) influenza virus. The anthropogenic housekeeping gene of beta-actin was selected and primers were designed as quality control for throat-swab samples. The reaction system of one-step four multiplex RT-PCR assay was established by optimizing the experimental conditions. The commercially available real-time fluorescent RT-PCR kits were used to validate this method.Results The one-step four multiplex RT-PCR assay for screening of 2013 novel avian influenza A(H7N9) virus was developed successfully.Conclusion The new method is simple, practical, low-cost and very suitable for rapid detection of 2013 novel avian influenza A(H7N9) virus.