Challenges for Precise Subtyping and Sequencing of a H5N1 Clade 2.3.4.4b Highly Pathogenic Avian Influenza Virus Isolated in Japan in the 2022-2023 Season Using Classical Serological and Molecular Methods

被引:1
作者
Komu, James G. [1 ,2 ]
Nguyen, Hiep Dinh [1 ]
Takeda, Yohei [3 ,4 ]
Fukumoto, Shinya [3 ,5 ]
Imai, Kunitoshi [3 ]
Takemae, Hitoshi [6 ]
Mizutani, Tetsuya [6 ]
Ogawa, Haruko [3 ]
机构
[1] Obihiro Univ Agr & Vet Med, Grad Sch Anim & Vet Sci & Agr, 2 11 Inada, Obihiro, Hokkaido 0808555, Japan
[2] Jomo Kenyatta Univ Agr & Technol, Dept Med Lab Sci, Coll Hlth Sci, POB 62000 00200, Nairobi, Kenya
[3] Obihiro Univ Agr & Vet Med, Dept Vet Med, 2 11 Inada, Obihiro, Hokkaido 0808555, Japan
[4] Obihiro Univ Agr & Vet Med, Res Ctr Global Agromed, 2 11 Inada, Obihiro, Hokkaido 0808555, Japan
[5] Obihiro Univ Agr & Vet Med, Natl Res Ctr Protozoan Dis, 2 11 Inada, Obihiro, Hokkaido 0808555, Japan
[6] Tokyo Univ Agr & Technol, Ctr Infect Dis Epidemiol & Prevent Res CEPiR, Tokyo, 1838509, Japan
来源
VIRUSES-BASEL | 2023年 / 15卷 / 11期
关键词
highly pathogenic avian influenza; classical diagnostic methods; antigenic and genetic evolution; PCR; EVOLUTION; NEURAMINIDASE; DIAGNOSIS;
D O I
10.3390/v15112274
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The continuous evolution of H5Nx highly pathogenic avian influenza viruses (HPAIVs) is a major concern for accurate diagnosis. We encountered some challenges in subtyping and sequencing a recently isolated H5N1 HPAIV strain using classical diagnostic methods. Oropharyngeal, conjunctival, and cloacal swabs collected from a dead white-tailed eagle (Haliaeetus albicilla albicilla) were screened via real-time RT-PCR targeting the influenza A virus matrix (M) gene, followed by virus isolation. The hemagglutination inhibition test was applied in order to subtype and antigenically characterize the isolate using anti-A/duck/Hong Kong/820/80 (H5N3) reference serum or anti-H5N1 cross-clade monoclonal antibodies (mAbs). Sequencing using previously reported universal primers was attempted in order to analyze the full-length hemagglutinin (HA) gene. Oropharyngeal and conjunctival samples were positive for the M gene, and high hemagglutination titers were detected in inoculated eggs. However, its hemagglutination activity was not inhibited by the reference serum or mAbs. The antiserum to a recently isolated H5N1 clade 2.3.4.4b strain inhibited our isolate but not older strains. A homologous sequence in the previously reported forward primer and HA2 region in our isolate led to partial HA gene amplification. Finally, next-generation sequencing confirmed the isolate as H5N1 clade 2.3.4.4b HPAIV, with genetic similarity to H5N1 strains circulating in Japan since November 2021.
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页数:12
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