Development of multiplex rt-PCR assays for rapid detection and subtyping of influenza type A viruses from clinical specimens.

摘要: We developed multiplex RT-PCR assays that can detect and identify 12 hemagglutinin (H1-H12) 9 neuraminidase (N1-N9) subtypes are commonly isolated from avian, swine, human influenza A viruses. products with unique sizes characteristic of each subtype were amplified by RT-PCRs, sequence analysis amplicon was demonstrated to be specific for 24 reference The specificity further DNA or cDNA templates 7 viruses, 5 bacteria, 50 virus negative specimens. Furthermore, the could up 105 dilution viruses had an original infectivity titer 106 EID50/ml. Of 188 isolates, results agreed completely individual subtyping obtained isolations. methods efficiently detected mixed infections at least two different in one host. Therefore, these facilitate rapid accurate directly field