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

Qiu, Baofeng; Liu, Wujie; Peng, Daxin; Hu, Shun-Lin; Tang, Yinghua; Liu, Xiufan

doi:10.1016/j.jviromet.2008.10.001

Cited by 53 publications

(30 citation statements)

References 21 publications

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“…Therefore, calculation of the virus titer by the BehrensKärber method was not applicable for the results on the tracheal sample. The conjunctival sample was confirmed to contain the H5N1 virus by subtype specific RT-PCR using the H5 primer and the N1 primer reported by Lee et al [8] and Qiu et al [13], respectively ( Table 1). The virus isolated from the conjunctival swab was designated as A/whooper swan/Hamanaka/2011 (H5N1) (Ws/HN/11).…”

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confidence: 75%

H5N1 highly pathogenic avian influenza virus isolated from conjunctiva of a whooper swan with neurological signs

et al. 2012

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An H5N1 highly pathogenic avian influenza virus was isolated from conjunctiva of a whooper swan with neurological signs, which was captured during the latest H5N1 HPAI outbreak in Japan. The conjunctival swab contained a larger amount of the virus in comparison with the tracheal swab. This is the first report on H5N1 virus isolation from the conjunctiva of a wild bird, and the result may suggest the conjunctival swab to be a critical sample for H5N1 HPAIV detection in waterfowl. Phylogenetic analysis of the HA gene indicated that the virus falls into H5N1 clade 2.3.2.1.

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confidence: 75%

H5N1 highly pathogenic avian influenza virus isolated from conjunctiva of a whooper swan with neurological signs

et al. 2012

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“…The presence of AIV was checked by real-time RT-PCR for detection of the M gene [28]. The AIV isolates were subtyped for HA and NA by RT-PCR and confirmed by serological tests according to the methods described previously [29][30][31]. Three of the 10 H5N2 viruses were Not determined due to a stop codon in the exon H5N2 influenza viruses isolated from wild birds in Japan 3311 isolated in Hakuryo Pond during 2009 from wild birds migrating through Hokkaido [27].…”

Section: Viruses and Virus Identificationmentioning

confidence: 99%

Genetic characterization of H5N2 influenza viruses isolated from wild birds in Japan suggests multiple reassortment

et al. 2016

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Low-pathogenic avian influenza viruses (LPAIVs) of the H5 subtype can mutate to highly pathogenic forms, potentially destabilizing the poultry industry. Wild migratory birds are considered a natural reservoir of LPAIVs capable of dispersing both high-and low-pathogenic forms of the virus. Therefore, surveillance and characterization of AIV in wild birds are essential. Here, we report on the isolation and genetic characterization of 10 AIVs of the H5N2 subtype obtained through surveillance in Hokkaido, Japan, during 2009 and 2011. Fullgenome sequencing revealed that the H5 and N2 genes of these isolates are all closely related to each other, belonging to the Eurasian avian-like lineage, but they are unrelated to H5 highly pathogenic strains of clade 2.3.4.4. The internal genes of the isolates were found to be diverse, consistent with our hypothesis that these H5N2 strains have undergone multiple reassortment events. Even though all of the H5N2 isolates were characterized as LPAIV based on the amino acid sequences at the HA cleavage site, this analysis demonstrates a diverse pool of precursors that may seed future outbreaks in poultry and possible human transmissions, suggesting the need for high-quality surveillance.

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“…htm). AI virus was subtyped by reverse transcription polymerase chain reaction (RT-PCR) assays as described previously (Qiu et al, 2009;Tsukamoto et al, 2008) and confirmed by sequencing.…”

Section: Introductionmentioning

confidence: 99%

Molecular Characterization of an H5N3 Influenza Virus Isolated from Spot-Billed Duck

Lee¹,

Kwon²,

Sung³

2013

Korean Journal of Poultry Science

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Among the 16 hemagglutinin (HA) subtypes of avian influenza virus (AIV), only the H5 and H7 subtypes have caused highly pathogenic avian influenza (HPAI) in poultry. However, most H5 or H7 subtype viruses are categorized as low pathogenic avian influenza (LPAI). Some AIVs, including the H5 and H7 HPAI viruses, have shown the ability to infect humans directly. In this study, we describe the biological and molecular characterization of an H5N3 AIV (SBD/KR/KNU SYG06/06) isolated from spot-billed duck (Anas poecilorhyncha) in Korea. A phylogenetic analysis of the eight viral genes showed that the SBD/KR/KNU SYG06/06 isolate belongs to the Eurasian lineage and that the SBD/KR/KNU SYG06/06 isolate was clearly different from HPAI H5N1 strains, including human isolates and the Italian HPAI H5N2 strains. Additionally, no relationship was found between SBD/KR/KNU SYG06/06 and the Korean HPAI H5N1 isolates. The SBD/KR/ KNU SYG06/06 isolate had avian specific receptor binding site residues in the HA protein and the four C-terminal amino acids in the NS1 protein. The HA protein of the SBD/KR/KNU SYG06/06 isolate exhibited the typical LPAI motif at the cleavage site and this virus produced no cytopathic effects in MDCK cells without trypsin. Given these results, we suggest that the H5N3 AIV isolated from the spot-billed duck should be considered an LPAI virus and should have no pathogenic effect in humans.

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

Cited by 53 publications

References 21 publications

H5N1 highly pathogenic avian influenza virus isolated from conjunctiva of a whooper swan with neurological signs

H5N1 highly pathogenic avian influenza virus isolated from conjunctiva of a whooper swan with neurological signs

Genetic characterization of H5N2 influenza viruses isolated from wild birds in Japan suggests multiple reassortment

Molecular Characterization of an H5N3 Influenza Virus Isolated from Spot-Billed Duck

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