NGUYEN TT, Kwon HJ, Kim IH, Hong SM, et al. Multiplex nested RT-PCR for detecting avian influenza virus, infectious bronchitis virus and Newcastle disease virus. J Virol Methods. 2012 Dec 19. pii: S0166-0934(12)0
In this study, multiplex nested RT-PCR (mnRT-PCR) was applied to simultaneous detect multiplex PCR with the higher sensitivity of nested PCR that is required for avian influenza, infectious bronchitis and Newcastle disease virus using two steps of amplification. For the first PCR, primers that were specific for each virus were newly designed from the nucleoprotein gene of AIV, the nucleocapsid protein gene of IBV and the fusion protein gene of NDV to amplify products of 665, 386 and 236 nucleotides, respectively. The multiplex PCR step provides mass amplification using common primers, which increased markedly the sensitivity of the test. Non-specific reactions were not observed when other viruses and bacteria were used for evaluating the mnRT-PCR. As a field application, 172 samples were tested by RT-PCR and mnRT-PCR. Among these samples, the concordance rates for mnRT-PCR and the single conventional RT-PCR showed 98.9% (kappa=0.98) and 98.8% (kappa=0.96) similarity for IBV and AIV, respectively. As a result, it is recommended the multiplex nested PCR as an effective tool for detecting and studying the molecular epidemiology of various mixed infections of one or more of these viruses in poultry.
See Also:
Latest articles in those days:
- Outbreak of Highly Pathogenic Avian Influenza A(H5N1) Virus in Seals, St. Lawrence Estuary, Quebec, Canada 2 days ago
- Molecular characterization of influenza virus circulating in Nepal in the year 2019 2 days ago
- Seasonal antigenic prediction of influenza A H3N2 using machine learning 2 days ago
- Probable extinction of influenza B/Yamagata and its public health implications: a systematic literature review and assessment of global surveillance databases 2 days ago
- Code to reproduce analysis in Nguyen et al, Emergence and interstate spread of highly pathogenic avian influenza A (H5N1) in dairy cattle. 2 days ago
[Go Top] [Close Window]