云南省第4例人感染欧亚类禽H1N1亚型猪流感病毒分子流行病学调查

孙艳红; 陈瑶瑶; 周洁楠; 张美玲; 刘照生; 赵晓南

云南省第4例人感染欧亚类禽H1N1亚型猪流感病毒分子流行病学调查

云南省疾病预防控制中心，云南昆明　650506

基金项目: 云南省疾病预防控制中心科技计划项目（2023ZX005）

详细信息

作者简介:
孙艳红（1986～），女，云南昆明人，医学硕士，主管技师，主要从事急性传染病防制工作

通讯作者:
赵晓南，E-mail：20386586@qq.com

中图分类号: R373.1
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- 文章访问数: 21
- HTML全文浏览量: 17
- PDF下载量: 1
- 被引次数: 1
出版历程
- 收稿日期: 2024-12-12
- 网络出版日期: 2025-04-05

Molecular Epidemiological Investigation of the Fourth Human Case of Eurasian Avian-like H1N1 Swine Influenza Virus Infection in Yunnan Province

Yunnan Center for Disease Control and Prevention，Kunming Yunnan 650506，China

摘要

摘要: 目的对2022年云南省境内发现的1例（全省第4例）欧亚类禽猪流感病例进行分子流行病学调查，掌握其基因特征，揭示该新亚型欧亚类禽猪流感病毒对人类健康潜在影响。方法使用实时荧光定量PCR检测技术对病例的咽拭子标本、密切接触者和居住环境标本进行核酸检测。使用MDCK细胞进行阳性标本病毒分离，使用豚鼠血红细胞凝集实验和实时荧光定量RT-PCR方法对细胞培养物进行鉴定。通过Illumina Miseq二代基因测序平台进行全基因组测序，使用MEGA7.0软件构建系统发育树并分析其基因分子特征。结果成功分离云南省首例G5基因型欧亚类禽猪流感病毒，并获得该病毒全基因组序列。该病毒具有在哺乳动物的适应性或毒力、传播性增强的相关分子特征，与江苏分离的一株猪源毒株序列具有99.2%～99.7%的核苷酸一致性。该病毒在人群中传播的风险进一步增加。结论加强猪流感监测工作对于预防可感染人类的新型流感病毒亚型具有重要意义。
- 欧亚类禽H1NI猪流感 /
- 二代测序 /
- 系统发育分析 /
- 氨基酸特征
Abstract: Objective To conduct a unique and pioneering molecular epidemiological investigation of a case of Eurasian avian-like H1N1 swine influenza identified in Yunnan Province in 2022 （the fourth such case in the province） and to understand its genetic characteristics so as to reveal its potential impact on human health. Methods Real-time fluorescent quantitative PCR detection technology was used for the nucleic acid testing of the case's pharyngeal swab samples, close contacts, and environmental samples from the living area. Positive samples were subjected to virus isolation using MDCK cells. Cell cultures were authenticated using erythrocyte agglutination assay with guinea pig blood and real-time fluorescence quantitative RT-PCR. Whole genome sequencing was performed using the Illumina MiseqNext-generation sequencing platform, and a phylogenetic tree was constructed using MEGA 7.0 software to analyze the genetic molecular characteristics. Results The first G5 genotype Eurasian avian-like H1N1 swine influenza virus in Yunnan Province was successfully isolated, and the whole genome sequence of the virus was obtained. This virus possessed the molecular characteristics associated with increased adaptability, virulence, or transmissibility in mammals and had a nucleotide consistency of 99.2%～99.7% with a porcine strain isolated in Jiangsu province. These findings underscored the potential threat this virus poses to human health. Conclusion The study underscores the importance of further monitoring swine influenza in preventing new influenza virus subtypes that can infect humans.
- Eurasian avian-like H1N1 swine influenza virus /
- Next-generation sequencing /
- Phylogenetic analysis /
- Amino acid characterization

HTML全文

图 1 YN/01/22感染MDCK细胞的病变（cytopathic effects，CPE）情况（164x）

A：MDCK细胞对照；B：48 h CPE；C：72 h CPE。

Figure 1. Cytopathic effects of MDCK cells infected with YN/01/22（164x）

下载: 全尺寸图片幻灯片

图 2 EA H1N1 SIV分离株基因进化树

A：PB2基因系统发育树；B：HA基因系统发育树；C：M基因系统发育树；D：NA基因系统发育树；E：NP基因系统发育树；F：NS基因系统发育树；G：PA基因系统发育树；H：PB1基因系统发育树。

Figure 2. The genetic phylogenetic tree of EA H1N1 SIV isolate

下载: 全尺寸图片幻灯片

表 1 不同型别的欧亚类禽猪流感基因型分型依据

Table 1. Genotyping basis of different types of eurasian avian-like swine influenza genotypes

分离株	不同来源的基因片段
分离株	HA	NA	PB2	PB1	PA	NP	M	NS	基因型
A/Jiangsu/1/2011^a	EA	EA	EA	EA	EA	EA	EA	EA	1
A/swine/Hong_Kong/72/2007^a	EA	EA	EA	EA	EA	EA	EA	CS	2
A/Hunan/42443/2015^a	EA	EA	PDM	PDM	PDM	PDM	EA	CS	3
A/swine/Guangdong/1361/2010^a	EA	EA	PDM	PDM	PDM	PDM	PDM	PDM	4
A/Fujian-cangshan/SWL624/2016^a	EA	EA	PDM	PDM	PDM	PDM	PDM	CS	5
A/swine/Guangdong/1/2010^a	EA	EA	CS	CS	CS	CS	EA	CS	6
A/swine/Hong_Kong/2421/2012^a	EA	EA	EA	EA	EA	EA	PDM	EA	7
A/swine/Guangdong/NS2897/2012^a	EA	EA	PDM	EA	PDM	PDM	PDM	EA	8
A/swine/Guangxi/S2/2013^a	EA	EA	EA	EA	PDM	PDM	EA	CS	9
A/swine/Hong_Kong/268/2012^a	EA	EA	CS	CS	CS	CS	CS	CS	10
A/swine/Hong_Kong/201/2010^a	EA	PDM	CS	CS	CS	CS	CS	CS	11
A/Yunnan-Lincang/ASWL01/2022^b	EA	EA	PDM	PDM	PDM	PDM	PDM	CS	5
a：欧亚禽猪流感基因型分型参考株；b：云南临沧人感染欧亚禽猪流感分离株；EA：基因片段来源于欧亚类禽猪流感病毒；CS：基因片段来源于经典猪流感病毒；PDM：基因片段来源于A（H1N1）pdm09流感病毒。

下载: 导出CSV

表 2 A/Yunnan-Lincang/ASWL01/2022毒株的氨基酸分子特征分析

Table 2. The amino acid molecular characteristics of A/Yunnan-Lincang/ASWL01/2022 strain

基因	氨基酸突变位点	意义	毒株
基因	氨基酸突变位点	意义	CA/07/09	HN/42443/15	GD/1/10	FJ/624/16	JS/HD11/20	YN/01/22
HA	E190D G225D/E	病毒受体结合特异性从α-2， 3-唾液酸转变为α-2，6-唾液酸^[20]	D D	D E	V E	D E	D E	D E
NA	H275Y^a N295S^a	神经氨酸酶抑制剂耐药位点	H N	H N	H N	H N	H N	H N
PB2	L89V T271A G590S Q591R D701N	被证明对增加聚合酶活性、增强病毒复制和传播性至关重要。	V A S R D	V A G R D	V A S R D	V A S Q D	V A S R D	V A S R D
	T588I	除增加哺乳动物细胞中的聚合酶活性外，还被证明介导干扰素-β表达的抑制	T	I	T	T	I	I
	627	宿主特异性的决定因素，禽流感是E，人类流感是K	E	E	E	E	E	E
NS1	P42S D92E	增加哺乳动物病毒毒力或适应性	S D	S D	S D	S D	S D	S D
M2	S31N	金刚烷胺类抗病毒药物耐药	N	N	N	N	N	N
M1	T215A	增加哺乳动物病毒毒力或适应性	A	A	A	A	A	A
PA	L336M K356R S409N	增加聚合酶活性、增强病毒复制和传播性至关重要	M R N	M R N	L K N	M R N	M R N	M R N
PB1	X99H I368R	物种之间传播	H I	H I	H I	H I	H I	H I
NP	Q357K	增加哺乳动物病毒毒力或适应性	K	K	K	K	K	K
a：H1 Numbering。

下载: 导出CSV

表 3 中国人感染EA H1N1SIV病例

Table 3. The cases infected EA H1N1SIV in China

年份	省份	毒株名	基因型	参考文献
2011	江苏	A/Jiangsu/1/2011	1	Yang et al.^[10]
2012	河北	A/Hebei-Yuhua/SWL1250/2012	1	Wang et al.^[11]
2015	湖南	A/Hunan/42443/2015	3	Zhu et al.^[13]
2015	云南	A/Yunnan-Longyang/SWL1982/2015	3	Zhu et al.^[14]
2015	云南	A/Yunnan-Wuhua/SWL1869/2015	3	Zhu et al.^[14]
2016	福建	A/Fujian-Cangshan/SWL624/2016	5	Xie et al.^[12]
2018	天津	A/Tianjin-baodi/1606/2018（H1N1）	5	Li et al.^[15]
2020	云南	A/Yunnan⁃Mengzi/1462/2020	4	Li，et al.^[17]
2022	云南	A/Yunnan-Lincang/ASWL01/2022	5	孙艳红，等

下载: 导出CSV

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