欧亚类禽 H1N1 猪流感重组病毒的遗传特征与致病性
Genetic characterization and pathogenicity of a Eurasian avian-like H1N1 swine influenza reassortant virus.
机构信息
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, Hubei, China.
Key Laboratory of Preventive Veterinary Medicine in Hubei Province, The Cooperative Innovation Center for Sustainable Pig Production, Wuhan, 430070, Hubei, China.
出版信息
Virol J. 2022 Dec 2;19(1):205. doi: 10.1186/s12985-022-01936-6.
BACKGROUND
Swine influenza viruses (SIV), considered the "mixing vessels" of influenza viruses, posed a significant threat to global health systems and are dangerous pathogens. Eurasian avian-like H1N1(EA-H1N1) viruses have become predominant in swine populations in China since 2016.
METHODS
Lung tissue samples were obtained from pregnant sows with miscarriage and respiratory disease in Heilongjiang province, and pathogens were detected by Next-generation sequencing (NGS) and PCR. The nucleic acid of isolates was extracted to detect SIV by RT-PCR. Then, SIV-positive samples were inoculated into embryonated chicken eggs. After successive generations, the isolates were identified by RT-PCR, IFA, WB and TEM. The genetic evolution and pathogenicity to mice of A/swine/Heilongjiang/GN/2020 were analyzed.
RESULTS
The major pathogens were influenza virus (31%), Simbu orthobunyavirus (15%) and Jingmen tick virus (8%) by NGS, while the pathogen that can cause miscarriage and respiratory disease was influenza virus. The SIV(A/swine/Heilongjiang/GN/2020) with hemagglutination activity was isolated from lung samples and was successfully identified by RT-PCR, IFA, WB and TEM. Homology and phylogenetic analysis showed that A/swine/Heilongjiang/GN/2020 is most closely related to A/swine/Henan/SN/10/2018 and belonged to EA-H1N1. Pathogenicity in mice showed that the EA-H1N1 could cause lethal or exhibit extrapulmonary virus spread and cause severe damage to respiratory tracts effectively proliferating in lung and trachea.
CONCLUSION
A/swine/Heilongjiang/GN/2020 (EA-H1N1) virus was isolated from pregnant sows with miscarriage and respiratory disease in Heilongjiang province, China. Clinical signs associated with influenza infection were observed during 14 days with A/swine/Heilongjiang/GN/2020 infected mice. These data suggest that A/swine/Heilongjiang/GN/2020 (EA-H1N1) had high pathogenicity and could be systemic spread in mice.
背景
猪流感病毒（SIV）被认为是流感病毒的“混合容器”，对全球卫生系统构成重大威胁，是危险的病原体。自 2016 年以来，欧亚类禽流感 H1N1（EA-H1N1）病毒已成为中国猪群中的主要流行病毒。
方法
从黑龙江省流产和呼吸道疾病的妊娠母猪中获取肺组织样本，通过下一代测序（NGS）和 PCR 检测病原体。提取分离物的核酸，通过 RT-PCR 检测 SIV。然后，将 SIV 阳性样本接种到鸡胚中。经过连续几代，通过 RT-PCR、IFA、WB 和 TEM 鉴定分离物。分析 A/swine/Heilongjiang/GN/2020 的遗传进化和对小鼠的致病性。
结果
NGS 检测的主要病原体为流感病毒（31%）、Simbu 正布尼亚病毒（15%）和京门 tick 病毒（8%），而引起流产和呼吸道疾病的病原体为流感病毒。从肺组织样本中分离出具有血凝活性的 SIV（A/swine/Heilongjiang/GN/2020），并通过 RT-PCR、IFA、WB 和 TEM 成功鉴定。同源性和系统发育分析表明，A/swine/Heilongjiang/GN/2020 与 A/swine/Henan/SN/10/2018 最为密切相关，属于 EA-H1N1。在小鼠中的致病性表明，EA-H1N1 可引起致死性或表现出肺外病毒传播，有效增殖于肺部和气管中，导致呼吸道严重损伤。
结论
从中国黑龙江省流产和呼吸道疾病的妊娠母猪中分离出 A/swine/Heilongjiang/GN/2020（EA-H1N1）病毒。感染 A/swine/Heilongjiang/GN/2020 的小鼠在 14 天内出现与流感感染相关的临床症状。这些数据表明，A/swine/Heilongjiang/GN/2020（EA-H1N1）具有高致病性，可在小鼠中全身传播。
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