2015-2017年盐城市乙型流感病毒HA1、NA基因分子特征

陈国清; 李春香; 王瑶; 李峰; 徐士林; 李长城; 邵荣标

doi:10.16462/j.cnki.zhjbkz.2020.02.005

2015-2017年盐城市乙型流感病毒HA1、NA基因分子特征

doi: 10.16462/j.cnki.zhjbkz.2020.02.005

1.
224002 盐城, 盐城市疾病预防控制中心病原微生物检验科
2.
224002 盐城市妇幼保健院遗传室

陈国清、李春香为共同第一作者

基金项目: 江苏省卫计委医学青年科研课题(Q201513);盐城市医学科技发展计划项目(YK2016022)

详细信息

通讯作者:
邵荣标, E-mail:604506383@qq.com

中图分类号: R181.2
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出版历程
- 收稿日期: 2019-08-22
- 修回日期: 2019-11-24
- 刊出日期: 2020-02-10

Molecular characteristics of the hemagglutinin 1 and neuraminidase genes of influenza B viruses isolated in Yancheng city from 2015 to 2017

1.
Laboratory of Pathogenic Microorganism, Yancheng Center for Disease Control and Prevention, Yancheng 224002 China
2.
Laboratory of Medical Genetics y Yancheng Maternity and Child Health Care Hospital, Yancheng 224002, China

CHEN Guo-qing and LI Chun-xiang contributed equally to this article

Funds: Scientific Research Project of Medical Youth in Jiangsu Provincial Health and Family Planning Commission(Q201513); Yancheng City Medical Science and Technology Development Program Project(YK2016022)

More Information

Corresponding author: SHAO Rong-biao, E-mail:604506383@qq.com

摘要

摘要: 目的对盐城市2015-2017年流行的乙型流感病毒血凝素(hemagglutinin, HA)和神经氨酸酶(neuraminidase, NA)基因进行分子进化特征研究。方法将盐城市2015-2017年流感监测哨点医院以及流感暴发点采集的流感样病例咽拭子标本进行核酸、病毒分离检测定型, 对选取的18株乙型流感病毒分离株采用一步法RT-PCR方法扩增其HA1基因和NA基因, 扩增产物经纯化测序, 采用生物信息软件从核苷酸、氨基酸以及分子进化层面对毒株进行分子特征分析。结果盐城市2015-2017年分离的乙型流感病毒HA1基因和NA基因聚类的分支关系基本一致, 2015年Yamagata系毒株位于Yamagata Clade 3分支, 属Phuket/3073类毒株; 2016-2017年Victoria系毒株分布于Victoria Clade 1A分支, 属Brisbane/60类毒株。Yamagata系所有毒株在190-helix抗原表位均发生了D196N位点的变异; Victoria系毒株共涉及2个抗原表位, 120-loop抗原表位的117、129位点, 190-helix抗原表位的197、199位点。盐城株未发生系内、系间重配。18株分离株未发生NA蛋白酶活性位点以及耐药位点的变化。结论 2015年Yamagata系毒株与疫苗株B/Phuket/3073/2013匹配性较好。2016-2017年Victoria系毒株HA1和NA抗原基因变异位点在积累, 这些变异位点的积累很可能会导致流感病毒发生实质性的抗原性的漂移, 降低与流感疫苗株的匹配度, 减弱流感疫苗的保护作用。
- 乙型流感 /
- Yamagata系 /
- Victoria系 /
- RT-PCR /
- HA基因 /
- NA基因 /
- 序列分析
Abstract: Objective To analyze the genetic characteristics of the hemagglutinin(HA) and neuraminidase(NA) genes of influenza B viruses isolated in Yancheng City from 2015 to 2017. Methods The throat swab specimens of influenza-like illness(ILI) from sentinel surveillance hospital and outbreak sites were collected and sent to Yancheng CDC for virus nucleic acids and virus isolation testing. After validation with serological tests, eighteen strains of influenza B virus isolates were selected to amplify their HA1 and NA genes through RT-PCR assay. Their molecular characteristics of the obtained viral HA1 and NA gene sequences were analyzed using bioinformation software from three aspects, including nucleic acid level, amino acid level and molecular evolution level. Results Basically, the clustering relationships and the branche patterns between HA1 and NA genes from the 18 Yancheng influenza B virus strains were similar. The Yamagata lineage strains in 2015 were distributed in the Yamagata Clade 3 branch, belonging to Phuket/3073 strains. The Victoria lineage strains in 2016-2017 were distributed in the Victoria Clade 1 A branch, belonging to Brisbane/60 strains. D196 N substitution was detected on HA1 protein in all of Yamagata lineage strains at 190-helix epitope; Amino acid substitutions of victoria lineage strains involved two antigenic epitopes, 117 and 129 sites of 120-loop epitope and 197 and 199 sites of 190-helix epitope. No Intra-lineage or inter-lineage rearrangements occurred in Yancheng strains. Eighteen influenza B strains had no mutations in catalytic residues and drug resistant sites of NA genes. Conclusion The Yamagata strains well matched with vaccine strain B/Phuket/3073/2013. The HA1 and NA genes of victoria lineage strains circulated in Yancheng City during 2016 to 2017 are changing gradually. The accumulation of these mutations will result in antigenic drift of victoria lineage strains and increase the mismatch of the IFV field stains with the available vaccine strains, which may reduce the protective effect of flu vaccine.
- Influenza B virus /
- Yamagata lineage /
- Victoria lineage /
- RT-PCR /
- Hemagglutinin gene /
- Neuraminidase gene /
- sequencing

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图 1 盐城市2015-2017年乙型流感病毒HA1基因(A)、NA基因(B)分子进化树

注:●表示疫苗株; ▲表示2015-2017年盐城市分离株; ◆表示2017年盐城市流感暴发株; ■表示江苏代表株

Figure 1. Phylogenetic tree of the HA1 and HA genes sequences of influenza B viruses in Yancheng City from 2015 to 2017

下载: 全尺寸图片幻灯片

表 1 盐城市2015-2017年代表株与疫苗株HA1、NA基因核苷酸和氨基酸同源性分析

Table 1. Analysis of vaccine strains of nucleotide and amino acid homology of HA1 and NA genes with Yancheng strains in 2015 to 2017

代表株基因	基因长度(bp)	核苷酸同源性(%)	氨基酸同源性(%)	参考株
4株代表株HA1基因	981	99.4~99.5	99.1	B/Phuket/3073/2013(B-Yam)
NA基因	1401	99.6~99.7	99.1~99.3
14株代表株HA1基因	981	98.1~98.5	98.2~99.4	B/Brisbane/60/2008(B-Vic)
NA基因	1401	98.7~99.1	96.7~97.6

下载: 导出CSV

表 2 2016-2017年盐城市分离的乙型Victoria系毒株HA1区氨基酸变异位点分析

Table 2. Analysis of variation in the amino acid sites of HA1 genes of influenza B viruses isolated in Yancheng City from 2016 to 2017

毒株	HA1氨基酸变异位点
毒株	75	76	117	129	189	197	199	221
B/Brisbane/60/2008	K	I	I	N	T	S	T	T
B/Tinghu/1368/2016	-	-	-	D	A	N	-	-
B/Tinghu/1450/2016	-	-	-	D	A	N	-	-
B/Tinghu/1456/2016	-	-	V	D	-	N	-	I
B/Tinghu/1822/2016	-	-	V	D	-	N	A	I
B/Tinghu/1814/2016	-	-	V	D	-	N	I	-
B/Tinghu/1918/2016	-	-	V	D	-	N	A	-
B/Tinghu/1348/2017	-	V	V	D	-	N	I	I
B/Tinghu/1519/2017	-	-	V	D	-	-	-	-
B/Tinghu/1654/2017	-	-	V	D	-	N	-	-
B/Tinghu/1665/2017	-	-	V		-	N	I	-
B/Tinghu/1698/2017	-	-	V	D	-	N	A	I
B/Tinghu/11655/2017	-	-	-	D	A	N	I	-
B/Tinghu/11700/2017	-	-	V	D	-	N	-	-
B/Sheyang/32/2017	E	-	V	D	-	N	-	-

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