2016—2020年安徽省手足口病相关柯萨奇病毒A组16型分子分型研究

doi:10.3969/j.issn.1672-2302.2023.04.005

热带病与寄生虫学 ›› 2023, Vol. 21 ›› Issue (4): 199-203.doi: 10.3969/j.issn.1672-2302.2023.04.005

2016—2020年安徽省手足口病相关柯萨奇病毒A组16型分子分型研究

冯明子¹(), 傅荔艳¹, 葛盈露², 马婉婉², 王鹏², 孙永², 史永林¹^,²()

1.安徽医科大学公共卫生学院，安徽合肥 230032
2.安徽省疾病预防控制中心，安徽省公共卫生研究院，安徽省医疗卫生重点专科病原微生物分子生物学实验室

收稿日期:2023-04-28 出版日期:2023-08-20 发布日期:2023-08-23
通信作者: 史永林 E-mail:931839973@qq.com;sylin540@126.com
作者简介:冯明子，女，硕士在读，研究方向：公共卫生检验与检疫。E-mail: 931839973@qq.com
基金资助:
安徽省自然科学基金项目(1708085QH186)

Molecular typing of coxsackievirus A16 associated hand, foot and mouth disease in Anhui Province, 2016-2020

FENG Mingzi¹(), FU Liyan¹, GE Yinglu², MA Wanwan², WANG Peng², SUN Yong², SHI Yonglin¹^,²()

1. School of Public Health, Anhui Medical University, Hefei 230032, Anhui Province, China
2. Anhui Provincial Center for Disease Control and Prevention; Public Health Research Institute of Anhui Province; Key Laboratory for Medical and Health Science of Anhui Province

Received:2023-04-28 Online:2023-08-20 Published:2023-08-23
Contact: SHI Yonglin E-mail:931839973@qq.com;sylin540@126.com

摘要/Abstract

摘要：

目的了解安徽省手足口病（hand, foot and mouth disease, HFMD）相关柯萨奇病毒A组16型（coxsackievirus A16, CV-A16）基因型别分布情况。方法通过中国疾病预防控制信息系统传染病报告管理信息系统收集安徽省2016—2020年HFMD病例资料，并收集HFMD患儿咽拭子标本。对荧光定量PCR检出的CV-A16核酸阳性病例样本采用RT-PCR扩增VP1区基因片段并测序。运用肠道病毒在线分型工具确定基因型别，再利用MEGA 6.0生物软件构建CV-A16 VP1区基因进化树并分析VP1区氨基酸变异位点。结果 2016—2020年安徽省累计报告HFMD实验室确诊病例16 650例，其中CV-A16确诊病例3 809例（占22.88%），各年中CV-A16确诊病例占比依次为20.72%（219/1 057）、22.91%（677/2 955）、23.89%（1 004/4 202）、42.58%（1 687/3 962）、4.96%（222/4 474）。对获取的436份CV-A16阳性标本进行测序，获得290条VP1区基因序列，均属于B基因型，其中B1a基因亚型 65条，B1b 225条。290条VP1区基因序列核苷酸同源性为86.80%~100.00%，与CV-A16原型株G-10核苷酸同源性为74.10%~76.80%。65条B1a毒株VP1区基因序列与B1a参考株核苷酸同源性为93.00%~99.90%，225条B1b毒株VP1区基因序列与B1b参考株核苷酸同源性为88.40%~99.60%。B1a基因亚型流行株在遗传进化上分为4个分支簇，B1b基因亚型流行株在遗传进化上分为6个分支簇。与原型株G-10 VP1区编码氨基酸位点变异分析显示，290条VP1区基因存在多个位点变异。结论 2016—2020年安徽省HFMD CV-A16流行株B1a与B1b基因亚型二者共同流行，应持续关注安徽省肠道病毒病原监测与分子进化变异情况。

关键词: 手足口病, 柯萨奇病毒A组16型, 分子分型, 安徽省

Abstract:

Objective To understand the genetic types of coxsackievirus A16 (CV-A16) associated with hand, foot and mouth disease (HFMD) in Anhui Province. Methods The case data on HFMD reported in Anhui Province from 2016 to 2020 were downloaded from the Infectious Disease Reporting and Management System under the China Disease Prevention and Control Information System, and the throat swab samples were collected from HFMD children during the corresponding period. The samples of positive CV-A16 nucleic acid detected by real-time PCR were selected, and amplified for VP1 gene using reverse transcription PCR. Then the positive PCR products were sequenced. Software Molecular Evolutionary Genetics Analysis (MEGA, version 6.0) was used to construct the phylogenetic tree and for analysis of the mutation at amino acid of VP1 gene. Results In total, 16 650 cases of HFMD confirmed by laboratory studies were reported in Anhui Province from 2016 to 2020, including 3 809 cases of CV-A16 types (22.88%). The annual constituent ratio of CV-A16 was 20.72% (219/1 057), 22.91% (677/2 955), 23.89% (1 004/4 202), 42.58% (1 687/3 962) and 4.96% (222/4 474), respectively in different years. 290 VP1 sequences were obtained from 436 CV-A16 positive specimens, all of which belonged to B genotype, including 225 from B1b and 65 from B1a. The nucleotide homology of 290 VP1 sequences was in between 86.80% and 100.00%, and that of G-10 VP1 sequence was in between 74.10% and 76.80%. The nucleotide homology between 65 strains VP1 of B1a and reference strains identified as genotype B1a in GenBank database was 93.00%-99.90%.The nucleotide homology between the VP1 sequence of 225 B1b strains and the reference strains identified as genotype B1b in GenBank database was 88.40%-99.60%. The B1a and B1b genotypes consisted of 4 sub-clusters and 6 sub-clusters in genetic evolution. Amino acid variation analysis of the prototype strain G-10 revealed mutations at many sites of CV-A16. Conclusion B1a and B1b gene subtypes of HFMD CV-A16 strains were co-prevalent in Anhui Province during 2016-2020. It is necessary to continually monitor the molecular evolution of enterovirus pathogens in Anhui Province.

Key words: Hand, foot and mouth disease, CV-A16, Molecular typing, Anhui Province

中图分类号:

R373.2

冯明子, 傅荔艳, 葛盈露, 马婉婉, 王鹏, 孙永, 史永林. 2016—2020年安徽省手足口病相关柯萨奇病毒A组16型分子分型研究[J]. 热带病与寄生虫学, 2023, 21(4): 199-203.

FENG Mingzi, FU Liyan, GE Yinglu, MA Wanwan, WANG Peng, SUN Yong, SHI Yonglin. Molecular typing of coxsackievirus A16 associated hand, foot and mouth disease in Anhui Province, 2016-2020[J]. Journal of Tropical Diseases and Parasitology, 2023, 21(4): 199-203.

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参考文献 16

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2016—2020年安徽省手足口病相关柯萨奇病毒A组16型分子分型研究

Molecular typing of coxsackievirus A16 associated hand, foot and mouth disease in Anhui Province, 2016-2020

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年份	VP1区基因序列数	B1a		B1b
年份	VP1区基因序列数	序列数	比例（%）	序列数	比例（%）
2016	66	4	6.06	62	93.94
2017	60	4	6.67	56	93.33
2018	64	6	9.38	58	90.62
2019	83	36	43.37	47	56.63
2020	17	15	88.24	2	11.76
合计	290	65	22.41	225	77.59

年份	基因（亚）型	序列数量	3	13	15	17	19	22	26	33	36	43	59	61	79	119	139	179	183	213	217	220
1951	A/G-10	1	G	T	R	G	A	S	E	N	E	L	Q	E	T	M	A	I	L	A	S	A
2016	B1a	4	P	N	Q	N	S	A	L	E	S	V	S	K	A	L	V	V	M	E	A	L
2017	B1a	4	P	N	Q	N	S	A	L	E	S	V	S	K	A	L	V	V	M	E	A	L
2018	B1a	6	P	N	Q	N	S	A	L	E	S	V	S	K	A	L	V	V	M	E	A	L
2019	B1a	36	P	N	Q	N	S	A/T	L	E	S	V	S	K	A	L	V	V	M	E	A	L
2020	B1a	15	P	N	Q	N	S	A/T	L	E	S	V	S	K	A	L	V	V	M	E	A	L
2016	B1b	62	P	N	Q	N	S/T	A	L	E	S	V/I	S/R	K	A	L	V	V	M	E	A	L
2017	B1b	56	P/S	N	Q	N	S	A/T	L	E	S	V/I	S	K	A	L	V	V	M	E	A	L
2018	B1b	58	P	N	Q	N	S/T	A	L	E	S	V/I	S	K	A	L	V	V	M	E	A	L
2019	B1b	47	P	N/S	Q	N	S	A	L	E	S	V/I	S	K/R	A	L	V	V	M	E	A	L
2020	B1b	2	P	N	Q	N	S	A	L	E	S	V	S	K	A	L	V	V	M	E	A	L

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