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Virol J. 2015 Apr 30;12(1):69. [Epub ahead of print]
Simultaneous detection of novel H7N9 and other influenza A viruses in poultry by multiplex real-time RT-PCR.
Xu X1, Bao H2, Ma Y3, Sun J4, Zhao Y5, Wang Y6, Shi J7, Zeng X8, Li Y9, Wang X10, Chen H11.
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Abstract
BACKGROUND:
A novel reassortant H7N9 influenza A virus has crossed the species barrier from poultry to cause human infections in China in 2013 and 2014. Rapid detection of the novel H7N9 virus is important to detect this virus in poultry and reduce the risk of an epidemic in birds or humans.
FINDINGS:
In this study, a multiplex real-time RT-PCR (rRT-PCR) assay for rapid detection of H7N9 and other influenza A viruses was developed. To evaluate the assay, various influenza A viruses, other avian respiratory viruses, and 1,070 samples from poultry were tested. Fluorescence signals corresponding to H7 and N9 subtypes were detected only when H7 and N9 subtypes were present, while the fluorescence signal for the influenza A M gene was detected in all specimens with influenza A strains. The fluorescent signal can be detected in dilutions as low as 56 copies per reaction for the H7, N9 and M genes. Intra- and inter-assay variability tests showed a reliable assay. In poultry samples, a comparison of rRT-PCR with virus isolation showed a high level of agreement.
CONCLUSIONS:
The multiplex rRT-PCR assay in this study has good specificity, sensitivity and reproducibility, and will be useful for laboratory surveillance and rapid diagnosis of H7N9 and other influenza A viruses.
PMID: 25925390 [PubMed - as supplied by publisher] Free full text
Simultaneous detection of novel H7N9 and other influenza A viruses in poultry by multiplex real-time RT-PCR.
Xu X1, Bao H2, Ma Y3, Sun J4, Zhao Y5, Wang Y6, Shi J7, Zeng X8, Li Y9, Wang X10, Chen H11.
Author information
Abstract
BACKGROUND:
A novel reassortant H7N9 influenza A virus has crossed the species barrier from poultry to cause human infections in China in 2013 and 2014. Rapid detection of the novel H7N9 virus is important to detect this virus in poultry and reduce the risk of an epidemic in birds or humans.
FINDINGS:
In this study, a multiplex real-time RT-PCR (rRT-PCR) assay for rapid detection of H7N9 and other influenza A viruses was developed. To evaluate the assay, various influenza A viruses, other avian respiratory viruses, and 1,070 samples from poultry were tested. Fluorescence signals corresponding to H7 and N9 subtypes were detected only when H7 and N9 subtypes were present, while the fluorescence signal for the influenza A M gene was detected in all specimens with influenza A strains. The fluorescent signal can be detected in dilutions as low as 56 copies per reaction for the H7, N9 and M genes. Intra- and inter-assay variability tests showed a reliable assay. In poultry samples, a comparison of rRT-PCR with virus isolation showed a high level of agreement.
CONCLUSIONS:
The multiplex rRT-PCR assay in this study has good specificity, sensitivity and reproducibility, and will be useful for laboratory surveillance and rapid diagnosis of H7N9 and other influenza A viruses.
PMID: 25925390 [PubMed - as supplied by publisher] Free full text