Detection of highly pathogenic zoonotic influenza virus H5N6 by reverse-transcriptase quantitative polymerase chain reaction

Virology Journal

Brian Meehan¹  Frank YK Wong¹  Chris Morrissy¹  Som Walker¹  Stacey Valdeter¹  Jianning Wang¹  Adam J Foord¹  Hans G Heine¹ 

[1] CSIRO Australian Animal Health Laboratory, 5 Portarlington Road, Geelong 3220, VIC, Australia

关键词: PCR;    Clade 2.3.4.4;    H5N6;    Molecular diagnosis;    Reassortant virus;    Avian influenza;    Influenza virus;   

Others  :  1131013
DOI  :  10.1186/s12985-015-0250-3

 received in 2014-10-08, accepted in 2015-01-28,  发布年份 2015

【 摘 要 】

Background

Variant high pathogenicity avian influenza (HPAI) H5 viruses have recently emerged as a result of reassortment of the H5 haemagglutinin (HA) gene with different neuraminidase (NA) genes, including NA1, NA2, NA5, NA6 and NA8. These viruses form a newly proposed HA clade 2.3.4.4 (previously provisionally referred to as clade 2.3.4.6), and have been implicated in disease outbreaks in poultry in China, South Korea, Laos, Japan and Vietnam and a human fatality in China. There is real concern that this new clade may be wide spread and not readily identified using existing diagnostic algorithms.

Findings

Fluorescent probe based reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR) assays were developed to facilitate the identification of novel clade 2.3.4.4 viruses of H5N6 subtype emerging in Asia. Assays were aimed at the haemagglutinin (HA) gene for clade identification and at the NA gene to identify N6. The HA assay employing a minor groove binder (MGB) probe was able to detect and differentiate A/duck/Laos/XBY004/2014(H5N6) and related influenza A(H5N6) virus isolates belonging to the proposed clade 2.3.4.4 from other H5 HPAI viruses. In addition, an Eurasian N6 assay was able to differentiate N6 from other NA subtypes.

Conclusions

Laos influenza A(H5N6) virus representative of proposed clade 2.3.4.4, was detected and differentiated from viruses in other H5N1 clades using a clade-specific HA RT-qPCR assay whereas the N6-NA subtype was determined by an Eurasian N6 RT-qPCR assay. Such a clade-specific assay would be of particular value for surveillance and in diagnostic laboratories where sequencing is not readily available.

【 授权许可】

   
2015 Heine et al.; licensee BioMed Central.

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