A reverse transcription-PCR for subtyping of the neuraminidase of avian influenza viruses

作者：Qiu Bao Feng; Liu Wu Jie; Peng Da Xin; Hu Shun Lin; Tang Ying Hua; Liu Xiu Fan^*

来源：Journal of Virological Methods, 2009, 155(2): 193-198.

DOI：10.1016/j.jviromet.2008.10.001

• Avian influenza virus
• RT-PCR
• NA-subtyping

To date, nine neuraminidase (NA) subtypes of avian influenza viruses have been identified. In order to differentiate the NA of avian influenza viruses rapidly, a reverse transcription PCR (RT-PCR) was developed. Nine pairs of NA-specific primers for the RT-PCR were designed based on the analysis of 509 complete NA sequences in GenBank. The primers were designed to amplify partial NA genes and each pair is unique to a single NA subtype (N1-N9). By nine RT-PCRs simultaneously in a set of separate tubes, the subtype of NA was determined by subsequent agarose gel electrophoresis and ethidium bromide staining, since only one of the nine RT-PCRs would give a product of expected size for each virus strain. In comparison with the established method of sequence analysis of 101 reference strains or isolates of avian influenza viruses, the RT-PCR method had a sensitivity of 97.3% and a specificity of 91.1% in subtyping avian influenza viruses. These results indicate that the RT-PCR method described below provides a specific and sensitive alternative to conventional NA-subtyping methods.

• 出版日期2009-2
• 单位扬州大学