Comparison of avian influenzavirus genetic detection assays and verification of real-time PCR results by electrophoresis
2015
Inoue, D. (Chuo Livestock Hygiene Service Center, Nagasaki Prefecture, Isahaya (Japan)) | Yoshino, F. | Mukohara, Y. | Iwamatsu, S.
The sensitivity of genetic detection assays of avian influenzavirus (AIV) was compared to show that real-time polymerase chain reaction (rPCR) was 10 to 10,000 times more sensitive than conventional PCR (cPCR). Nucleotide sequences of the primers and probes for AIV genetic detection assays were compared with influenzavirus sequences registered to GenBank. A few mismatches between the nucleotide sequences of the primers and probes for the assays targeting the NP gene and those of the strains were observed. Several mismatches were observed in a case study targeting H5 genes, raising concern that sensitivity might be lower against several low pathogenic strains. Eelectrophoresis of an rPCR product was used for evaluation of the results, showing that it is rapid and sensitive enough to discriminate non-specific reaction caused in the rPCR assay.
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