2021—2023年安徽省手足口病柯萨奇病毒A组16型分子分型研究

doi:10.20199/j.issn.1672-2302.2024.04.009

摘要/Abstract

摘要：

目的了解安徽省手足口病（hand, foot and mouth disease, HFMD）发病情况、柯萨奇病毒A组16型（coxsackievirus A16, CV-A16）基因型别分布情况和VP1基因进化特征。方法通过中国疾病预防控制信息系统对2021—2023年安徽省HFMD发病情况和病原学检测情况进行统计。收集HFMD患者咽拭子样本，分离得到CV-A16 毒株，对其VP1区进行PCR扩增和基因序列测定，使用肠道病毒在线分型工具鉴定基因亚型，运用MegAlign比较核苷酸和氨基酸同源性，利用MEGA 6.0构建进化树，并分析 VP1 区氨基酸变异位点。结果 2021—2023年安徽省累计报告HFMD病例155 640例，年均报告发病率为84.83/10万。2021—2023年安徽省16个地市共报告HFMD实验室确诊病例12 643例，其中CV-A16占18.16%（2 296/12 643），各年度构成比分别为25.81%（1 127/4 366）、33.69%（782/2 321）和6.50%（387/5 956）。2021—2023年安徽省共分离到HFMD CV-A16 毒株72株，其中B1a和B1b 基因亚型分别为58株（占80.56%）、14株（占19.44%）。两种亚型毒株与CV-A16原型株G-10的核苷酸和氨基酸同源性分别为73.80%~76.20%和90.60%~92.30%。进化树显示B1a基因亚型流行株在遗传进化上分为 4个分支簇，B1b基因亚型流行株形成3个分支簇。72条VP1区基因序列与原型株比较发现了26个氨基酸位点发生变异。结论 2021—2023年安徽省HFMD CV-A16主要流行的基因亚型为B1a和B1b，B1a为优势基因亚型，应加强对CV-A16流行毒株的分子监测。

关键词: 手足口病, 柯萨奇病毒A组16型, VP1编码区, 进化分析, 安徽省

Abstract:

Objective To understand the epidemic of hand, foot and mouth Disease (HFMD) in Anhui Province, and analyze the genetic types and the genetic evolution of the VP1 coding region of coxsackievirus A16 (CV-A16). Methods The data on the incidence of HFMD in Anhui Province were retrieved from the China Information System for Disease Prevention and Control from 2021 through 2023, and statistics was conducted regarding the incidence and composition of each enterovirus serotype of HFMD. The throat swab samples were collected from HFMD patients, and from which CV-A16 strains were isolated. Then the VP1 region of CV-A16 was amplified and sequenced. The genotype/sub genotype of CV-A16 strands was identified by using enterovirus online typing tool, MegAlign Pro sequence alignment software was used to compare the homology of nucleotides and amino acids. The phylogenetic tree of VP1 gene was constructed by biological software MEGA 6.0 Software, and the mutation at amino acid of VP1 gene was analyzed. Results A total of 155 640 HFMD cases were reported in Anhui Province from 2021 to 2023. The annual reported incidence was 84.83 /100 000. Between 2021 and 2023, a total of 12 643 laboratory confirmed cases of HFMD were reported in 16 cities in Anhui area, in which CV-A16 associated cases accounted for 18.16% (2 296/12 643). The annual constituent ratio of CV-A16 for each year was 25.81% (1 127/4 366), 33.69% (782/2 321) and 6.50% (387/5 956), respectively. Seventy-two CV-A16 strains were isolated from HFMD cases in Anhui Province from 2021 to 2023, in which 58 strains were associated with genetic subtypes of B1a and 14 from B1b, respectively accounting for 80.56% and 19.44%. The nucleotide and amino acid homology with the CV-A16 prototype strain G-10 was 73.80%-76.20% and 90.60%-92.30%. The B1a and B1b genotypes consisted of 4 sub-clusters and 3 sub-clusters in genetic evolution. A total of 72 VP1 gene sequences were compared with the prototype strain and 26 amino acid sites were mutated. Conclusion The main prevalent gene subtypes of HFMD CV-A16 in Anhui area from 2021 to 2023 were involved in B1a and B1b, and B1a was predominant. Our findings suggest that monitoring of CV-A16 epidemic strains should be strengthened in Anhui Province.

Key words: Hand, foot and mouth disease, Coxsackievirus A16, VP1 coding region, Phylogenetic analysis, Anhui Province

中图分类号:

R373.2

葛盈露, 杨灵康, 刘以诺, 马婉婉, 王鹏, 孙永, 史永林. 2021—2023年安徽省手足口病柯萨奇病毒A组16型分子分型研究[J]. 热带病与寄生虫学, 2024, 22(4): 233-238.

GE Yinglu, YANG Lingkang, LIU Yinuo, MA Wanwan, WANG Peng, SUN Yong, SHI Yonglin. Molecular characterization of coxsackievirus A16 associated hand, foot and mouth disease in Anhui Province from 2021 to 2023[J]. Journal of Tropical Diseases and Parasitology, 2024, 22(4): 233-238.

图/表 4

图1

表1

图2

表2

参考文献 27

[1]	Liu WY, Wu SM, Xiong Y, et al. Co-circulation and genomic recombination of coxsackievirus A16 and enterovirus 71 during a large outbreak of hand, foot, and mouth disease in Central China[J]. PLoS One, 2014, 9(4):e96051.
[2]	Esposito S, Principi N. Hand, foot and mouth disease: current knowledge on clinical manifestations, epidemiology, aetiology and prevention[J]. Eur J Clin Microbiol Infect Dis, 2018, 37(3):391-398.
[3]	Zhu Z, Zhu SL, Guo XB, et al. Retrospective seroepidemiology indicated that human enterovirus 71 and coxsackievirus A16 circulated wildly in central and Southern China before large-scale outbreaks from 2008[J]. Virol J, 2010,7:300.
[4]	Cabrerizo M, Tarragó D, Muñoz-Almagro C, et al. Molecular epidemiology of enterovirus 71, coxsackievirus A16 and A6 associated with hand, foot and mouth disease in Spain[J]. Clin Microbiol Infect, 2014, 20(3):O150-O156.
[5]	Hosoya M, Kawasaki Y, Sato M, et al. Genetic diversity of coxsackievirus A16 associated with hand, foot, and mouth disease epidemics in Japan from 1983 to 2003[J]. J Clin Microbiol, 2007, 45(1) :112-120. pmid: 17093028
[6]	Wang JY, Teng Z, Chu W, et al. The emergence and spread of one Coxsackievirus A16 Genogroup D novel recombinant strain that caused a clustering HFMD outbreak in Shanghai,China, 2016[J]. Emerg Microbes Infect, 2018, 7(1):131.
[7]	Hassel C, Mirand A, Farkas A, et al. Phylogeography of coxsackievirus A16 reveals global transmission pathways and recent emergence and spread of a recombinant genogroup[J]. J Virol, 2017, 91(18):e00630-e00617.
[8]	Zhang Y, Wang DY, Yan DM, et al. Molecular evidence of persistent epidemic and evolution of subgenotype B1 coxsackievirus A16-associated hand, foot, and mouth disease in China[J]. J Clin Microbiol, 2010, 48(2):619-622. doi: 10.1128/JCM.02338-09 pmid: 20018819
[9]	Han ZZ, Song Y, Xiao JB, et al. Genomic epidemiology of coxsackievirus A16 in mainland of China, 2000-18[J]. Virus Evol, 2020, 6(2):veaa084.
[10]	马婉婉, 龚磊, 肖永康, 等. 2015—2022年安徽省手足口病流行病学特征及病原谱变化[J]. 安徽医科大学学报, 2023, 58(10):1763-1768.
[11]	卫生部. 手足口病预防控制指南(2009版)[J]. 全科医学临床与教育, 2010, 8(2):125-127,133.
[12]	蔡靖靖, 栾荣生. 2017—2022年四川省法定传染病流行特征分析[J]. 现代预防医学, 2023, 55(9):1566-1571.
[13]	尼格德力·阿力腾赛尔, 张璇, 陈媛, 等. 2018—2022年新疆维吾尔自治区手足口病流行特征分析[J]. 热带病与寄生虫学, 2023, 21(4):191-194,215.
[14]	温莹, 罗日景, 程雁鹏, 等. 2013—2022年深圳市托幼儿童传染病流行特征及趋势分析[J]. 现代预防医学, 2024,(1):149-155.
[15]	国务院应对新型冠状病毒感染疫情联防联控机制综合组. 关于印发对新型冠状病毒感染实施“乙类乙管”总体方案的通知[A/OL].(2022-12-06)[2023-07-09]. http://www.nhc.gov.cn/xcs/zhengcwj/202212/e97e4c449d7a475794624b8ea12123c6.shtml.
[16]	张雨桐, 宋杨, 刘凤凤, 等. 新型冠状病毒感染“乙类乙管”后我国手足口病流行特征与趋势分析[J]. 热带病与寄生虫学, 2023, 21(4):186-190,227.
[17]	史永林, 王贤, 陈国平, 等. 2014年安徽省柯萨奇病毒A组16型分离株VP1基因特征研究[J]. 病毒学报, 2015, 31(6):660-664.
[18]	冯明子, 傅荔艳, 葛盈露, 等. 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.
[19]	史永林, 葛盈露, 马婉婉, 等. 2018—2019年安徽省柯萨奇病毒A组16型B1基因亚型分离株分子特征分析[J]. 疾病监测, 2022, 37(8):1027-1032.
[20]	安徽省疾病预防控制中心. 安徽省2018年手足口病疫情专题风险评估报告[J]. 安徽预防医学杂志, 2018, 24(3):156-159.
[21]	Yi LN, Zeng HR, Zheng HY, et al. Molecular surveillance of coxsackievirus A16 in Southern China, 2008-2019[J]. Arch Virol, 2021, 166(6):1653-1659. doi: 10.1007/s00705-021-05052-8 pmid: 33796884
[22]	黄京京, 韦欢欢, 陈飞, 等. 2018—2019年北京市柯萨奇病毒A组16型分子流行病学研究[J]. 微生物学免疫学进展, 2023, 51(1):40-48.
[23]	李希尚, 赵丽娟, 伏晓庆, 等. 云南省保山市2022年手足口病病例柯萨奇病毒A16型基因特征[J]. 中国热带医学, 2023, 23(10):1104-1108. doi: 10.13604/j.cnki.46-1064/r.2023.10.16
[24]	王珊, 焦伯延, 冯强, 等. 2021年济宁市柯萨奇病毒A16型流行特点及全基因组特征[J]. 山东大学学报(医学版), 2023, 61(5):90-95.
[25]	Hoa-Tran TN, Dao ATH, Nguyen AT, et al. Coxsackieviruses A6 and A16 associated with hand, foot, and mouth disease in Vietnam, 2008-2017: essential information for rational vaccine design[J]. Vaccine, 2020, 38(52):8273-8285. doi: 10.1016/j.vaccine.2020.11.031 pmid: 33223308
[26]	魏雷雷, 沈博, 赵庆龙, 等. 吉林省2018—2021年柯萨奇病毒A组16型病毒基因特征分析[J]. 中国卫生工程学, 2023, 22(3):329-331.
[27]	徐莉, 崔大伟, 戴玉柱, 等. 浙江省柯萨奇病毒A16 VP1基因特征分析及B细胞抗原表位预测[J]. 临床检验杂志, 2017, 35(1):65-71.

年份	VP1区基因序列数	B1a亚型		B1b亚型
年份	VP1区基因序列数	毒株数	占比（%）	毒株数	占比（%）
2021	24	18	75.00	6	25.00
2022	36	28	77.78	8	22.22
2023	12	12	100.00	0	0
合计	72	58	80.56	14	19.44

基因型/亚型	3	11	13	14	15	17	19	22	23	26	33	36	43	59	61	79	119	139	164	179	183	213	215	217	220	251
A/G-10	G	A	T	S	R	G	A	S	L	E	N	E	L	Q	E	T	M	A	T	I	L	A	P	S	A	V
2021-B1a（n=18）	P	T	N	N	Q	N	S	A/T	L	L	E	S	V	S	K	A	L	V	K	V	M	E	L	A/V	L	I
2022-B1a（n=28）	P	T	N	N	Q	N	S	A/T	L	L	E	S	V	S	K/R	A	L	V	K	V	M	E	L	A	L	I
2023-B1a（n=12）	P	T	N	N	Q	N	S	A/T	L	L	E	S	V	S	K/R	A	L	V	K	V	M	E	L	A	L	I
2021-B1b（n=6）	P	T	N	S	Q	N	S	A	M	L	E	S	V	S	K	A	L	V	T	V	M	E	L	A	L	V
2022-B1b（n=8）	P	T	N	S	Q	N	S	A	M	L	E	S	V/I	S	K	A	L	V	T	V	M	E	L	A	L	V