| Title | Development of Multiplex RT-PCR Assays for Rapid Detection and Subtyping of Influenza Type A Viruses from Clinical Specimens. | | Author(s) | Chang HK, Park JH, Song MS, Oh TK, Kim SY, Kim CJ, Kim H, Sung MH, Han HS, Hahn YS, Choi YK | | Institution | College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju 361-763, Korea. | | Source | J Microbiol Biotechnol 2008 Jun; 18(6):1164-1169. | | Abstract | We developed multiplex RT-PCR assays that can detect and identify 12 hemagglutinin (H1-H12) and 9 neuraminidase (N1-N9) subtypes that are commonly isolated from avian, swine, and human influenza A viruses. RT-PCR products with unique sizes characteristic of each subtype were amplified by multiplex RT-PCRs, and sequence analysis of each amplicon was demonstrated to be specific for each subtype with 24 reference viruses. The specificity was demonstrated further with DNA or cDNA templates from 7 viruses, 5 bacteria, and 50 influenza A virus negative specimens. Furthermore, the assays could detect and subtype up to 105 dilution of each of the reference viruses that had an original infectivity titer of 106 EID50/ml. Of 188 virus isolates, the multiplex RT-PCR results agreed completely with individual RT-PCR subtyping results and with results obtained from virus isolations. Furthermore, the multiplex RT-PCR methods efficiently detected mixed infections with at least two different subtypes of influenza viruses in one host. Therefore, these methods could facilitate rapid and accurate subtyping of influenza A viruses directly from field specimens. | | Language | ENG | | Pub Type(s) | JOURNAL ARTICLE
| | PubMed ID | 18600063 |
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