[Rapid detection of novel avian influenza virus subtype H7N9 by multiplex real-time RT-PCR].

In order to develop a rapid detection kit for novel avian influenza virus(AIV)subtype H7N9, two sets of specific primers and probes were designed based on the nucleotide sequences of hemagglutinin antigen(HA)and neuraminidase antigen(NA)of novel H7N9virus(2013)available in GenBank to establish the method of TaqMan probe-based multiplex real-time RT-PCR for rapid detection of AIV subtype H7N9.The primer and probe of HA were for all H7subtype AIVs,while the primer and probe of NA were only for novel N9subtype AIVs.The results showed that this method had high sensitivity and specificity.This method was applicable to the testing of positive standard sample with a minimum concentration of 10copies/μL;it not only distinguished H7subtype from H1,H3,H5,H6,and H9subtypes, but also distinguished novel N9subtype from traditional N9subtype.A total of 2700samples from Zhuhai,China were tested by this method,and the results were as expected.For the advantages of sensitivity and specificity,the method holds promise for wide application.