| 187 | 0 | 301 |
| 下载次数 | 被引频次 | 阅读次数 |
目的 本文对2022-2023年云南省昆明市手足口病（Hand, foot and mouth disease, HFMD）病例样本进行基因测序定型并对其中常见病原体柯萨奇病毒A组4型（Coxsackievirus A4,CVA4）、6型（CVA6）、10型（CVA10）和16型（CVA16）的VP1区基因特征进行分析。方法 对2022-2023年昆明市经实时荧光定量PCR法鉴定为EVA71、CVA16或通用型阳性的161份HFMD病例粪便样本先进行VP4/VP2结合区基因测序定型，确定病毒型别。再测定各型病毒VP1区完整序列，并按文献下载CVA4、CVA6、CVA10和CVA16型病毒VP1基因代表株序列，用Mega 6.0软件构建基因进化树，进行基因特征分析。结果 2022-2023年昆明市疾控中心将经实时荧光定量PCR法检测阳性的随机抽取样本161份（2022年52份，2023年109份）送云南省疾病预防控制中心手足口病实验室进行基因测序定型，所有样本均能被扩增并测序定型，测序定型率为100%(161/161)。病毒检出率较多的依次是CVA6(48株，占29.81%,48/161)、CVA4(39株，占24.22%,39/161)、CVA10(37株，占22.98%,37/161)、CVA16(23株，占14.30%,23/161)。其他分离较少的14株病毒依次是CVA2 1株，CVB5 7株，E3 2株，PV1 2株（疫苗株），PV3 2株（疫苗株）。对这4种病毒完整VP1区基因特征分析表明，48株CVA6全为D3a亚型，39株CVA4为C2亚型，37株CVA10为基因型C,23株CVA16为B1a亚型。结论2022-2023年昆明市手足口病病原谱主要为CVA6、CVA4、CVA10和CVA16，基因特征分析表明，昆明市流行的4种主要病毒的基因型或亚型与全国其他省份一致，表明这些病毒的特定基因型和亚型在中国较长时间存在和流行，加强昆明市HFMD病原学监测和分子流行病学研究具有重要意义。
Abstract：Objective To determine the pathogenic spectrum of hand, foot and mouth disease(HFMD) in Kunming, Yunnan Province, China,from 2022 to 2023, and to analyze the VP1 gene characteristics of common pathogens, including Coxsackievirus A4(CVA4), CVA6, CVA10, and CVA16. Methods A total of 161 stool samples from HFMD cases collected in Kunming during 2022-2023, which were positive for EV-A71, CVA16 or pan-enterovirus detected by real-time fluorescent quantitative PCR, were selected for molecular typing.. Viral genotypes were first determined by sequencing the VP4/VP2 junction region. Complete VP1 gene sequences were subsequently obtained for each viral type. Representative VP1 sequences of CVA4, CVA6, CVA10, and CVA16 were downloaded from published databases, and phylogenetic trees were constructed using MEGA version 6.0 to analyze genetic characteristics. Results All 161 samples were successfully amplified and genotyped, yielding a sequencing success rate of 100%(161/161). The most frequently detected viruses were CVA6(48 strains, 29.81%), CVA4(39 strains, 24.22%), CVA10(37 strains, 22.98%), and CVA16(23 strains, 14.30%), all of which belong to enterovirus species A. Fourteen additional strains were identified at lower frequencies, including CVA2(1 strain), CVB5(7 strains), echovirus 3(2 strains), poliovirus type 1(2 vaccine strains), and poliovirus type 3(2 vaccine strains). Genetic analysis of the complete VP1 region showed that all CVA6 strains belonged to subgenotype D3a, CVA4 strains to subgenotype C2, CVA10 strains to genotype C, and CVA16 strains to subgenotype B1a. Conclusion From 2022 to 2023, the predominant pathogens causing HFMD in Kunming were CVA6, CVA4, CVA10, and CVA16. The genotypes and subgenotypes of these major circulating viruses were consistent with those reported in other regions of China, suggesting long-term persistence and circulation of these viral lineages nationwide. These findings highlight the importance of continued etiological surveillance and molecular epidemiological studies of HFMD in Kunming.
[1]中华人民共和国卫生部.手足口病预防控制指南（2008年版）[J].首都公共卫生，2008, 2(4):146-148. DOI:10. 3969/j. issn. 1673-7830. 2008. 04. 002.
[2]Chen KT, Chang HL, Wang ST, et al. Epidemiologic features of hand-foot-mouth disease and herpangina caused by enterovirus 71 in Taiwan, 1998-2005[J].Pediatrics, 2007, 120(2):e244-e252. DOI:10. 1542/peds. 2006-3331.
[3]Yu W, Xu H, Yin C. Molecular epidemiology of human coxsackievirus A16 strains[J]. Biomed Rep,2016, 4(6):761-764. DOI:10. 3892/br. 2016. 663.
[4]Xu M, Su L, Cao L, et al. Genotypes of the enterovirus causing hand foot and mouth disease in Shanghai, China, 2012-2013[J]. PLoS One, 2015, 10(9):e0138514. DOI:10. 1371/journal. pone. 0138514.
[5]Guo WP, Lin XD, Chen YP, et al. Fourteen types of co-circulating recombinant enterovirus were associated with hand, foot, and mouth disease in children from Wenzhou, China[J]. J Clin Virol, 2015, 70:29-38.DOI:10. 1016/j. jcv. 2015. 06. 093.
[6]Puenpa J, Mauleekoonphairoj J, Linsuwanon P, et al.Prevalence and characterization of enterovirus infections among pediatric patients with hand foot mouth disease,herpangina and influenza like illness in Thailand, 2012[J]. PLoS One, 2014, 9(6):e98888. DOI:10. 1371/journal. pone. 0098888.
[7]Fujimoto T, Iizuka S, Enomoto M, et al. Hand, foot,and mouth disease caused by coxsackievirus A6, Japan,2011[J]. Emerg Infect Dis, 2012, 18(2):337-339.DOI:10. 3201/eid1802. 111147.
[8]Lu QB, Zhang XA, Wo Y, et al. Circulation of coxsackievirus A10 and A6 in hand-foot-mouth disease in China, 2009–2011[J]. PLoS One, 2012, 7(12):e52073. DOI:10. 1371/journal. pone. 0052073.
[9]Zhang Y, Zhu Z, Yang W, et al. An emerging recombinant human enterovirus 71 responsible for the2008 outbreak of hand foot and mouth disease in Fuyang city of China[J]. Virol J, 2010, 7:94. DOI:10. 1186/1743-422X-7-94.
[10]Xing W, Liao Q, Viboud C, et al. Hand, foot, and mouth disease in China, 2008–12:an epidemiological study[J]. Lancet Infect Dis, 2014, 14(4):308-318.DOI:10. 1016/S1473-3099(13)70342-6.
[11]Xu Q, Qiu X, Yu Y, Wen Y, Zhang G, Li J.Epidemiology and genetic characteristics of coxsackievirus A16 associated with hand-foot-mouth disease in Hangzhou city, Zhejiang province from 2021to 2024. Front Microbiol. 2025 Nov 14;16:1698485.DOI:10. 3389/fmicb. 2025. 1698485..
[12]国家脊髓灰质炎和国家麻疹实验室.手足口病实验室手册[M]. 2010年第4版.北京：中国疾病预防控制中心，2010年：1-45.
[13]Ishiko H, Shimada Y, Yonaha M, et al. Molecular diagnosis of human enteroviruses by phylogeny-based classification by use of the VP4 sequence[J]. J Infect Dis, 2002, 185(6):744-754. DOI:10. 1086/339298.
[14]Xu Q, Qiu X, Yu Y, Wen Y, Zhang G, Li J.Epidemiology and genetic characteristics of coxsackievirus A16 associated with hand-foot-mouth disease in Hangzhou city, Zhejiang province from 2021to 2024. Front Microbiol. 2025 Nov 14;16:1698485.DOI:10. 3389/fmicb. 2025. 1698485.
[15]Oberste MS, Maher K, Williams AJ, et al. Speciesspecific RT-PCR amplification of human enteroviruses:a tool for rapid species identification of uncharacterized enteroviruses[J]. J Gen Virol, 2006, 87(Pt 1):119-128. DOI:10. 1099/vir. 0. 81179-0.
[16]Oberste MS, Maher K, Kilpatrick DR, et al. Typing of human enteroviruses by partial sequencing of VP1[J]. J Clin Microbiol, 1999, 37(5):1288-1293. DOI:10. 1128/JCM. 37. 5. 1288-1293. 1999.
[17]Oberste MS, Maher K, Kilpatrick DR, et al. Molecular evolution of the human enteroviruses:correlation of serotype with VP1 sequence and application to picornavirus classification[J]. J Virol, 1999, 73(3):1941-1948. DOI:10. 1128/JVI. 73. 3. 1941-1948. 1999.
[18]Oberste MS, Michele SM, Maher K, et al. Molecular identification and characterization of two proposed new enterovirus serotypes, EV74 and EV75[J]. J Gen Virol, 2004, 85(Pt 11):3205-3212. DOI:10. 1099/vir. 0. 80148-0.
[19]Ji T, Guo Y, Lv L, et al. Emerging recombination of the C2 sub-genotype of HFMD-associated CV-A4 is persistently and extensively circulating in China[J]. Sci Rep, 2019, 9(1):13668. DOI:10. 1038/s41598-019-49859-7.
[20]Song Y, Zhang Y, Ji T, et al. Persistent circulation of Coxsackievirus A6 of genotype D3 in mainland of China between 2008 and 2015[J]. Sci Rep, 2017, 7(1):5491. DOI:10. 1038/s41598-017-05618-0.
[21]Tian H, Zhang Y, Sun Q, et al. Prevalence of multiple enteroviruses associated with hand, foot, and mouth disease in Shijiazhuang City, Hebei Province, China:outbreaks of coxsackieviruses a10 and b3[J]. PLoS One, 2014, 9(1):e84233. DOI:10. 1371/journal.pone. 0084233.
[22]Zhang Y, Wang D, Yan D, et al. Molecular evidence of persistent epidemic and evolution of subgenotype B1coxsackievirus A16-associated hand, foot, and mouth disease in China[J]. J Clin Microbiol, 2010, 48(2):619-622. DOI:10. 1128/JCM. 02338-09.
[23]Guan H, Wang J, Wang C, et al. Etiology of multiple non-EV71 and non-CVA16 enteroviruses associated with hand, foot and mouth disease in Jinan, China, 2009-June 2013[J]. PLoS One, 2015, 10(11):e0142733.DOI:10. 1371/journal. pone. 0142733.
[24]Li J, Sun Y, Du Y, et al. Characterization of coxsackievirus A6-and enterovirus 71-associated hand foot and mouth disease in Beijing, China, from 2013 to2015[J]. Front Microbiol, 2016, 7:391. DOI:10. 3389/fmicb. 2016. 00391.
[25]张瀚文，付艺亮，李飞，等. 2019-2024年北京地区儿童手足口病病原学监测及流行病学分析[J].病毒学报，2025,41(06):1702-1709. DOI:10. 13242/j. cnki.bingduxuebao. 250228.
[26]尉秀霞，肖金波，李敏，等.聚集性发热疫情相关的柯萨奇病毒A组4型VP1区基因特征分析[J].病毒学报，2025,41(05):1361-1366. DOI:10. 13242/j. cnki.bingduxuebao. 250117.
[27]潘虹，陈小玲，文奇，等. 2018-2023年新余地区手足口病流行特征及其CVA6型病毒分子特征研究[J].病毒学报，2025,41(03):645-651. DOI:10. 13242/j. cnki.bingduxuebao. 240217.
[28]Mao QY, Wang Y, Bian L, et al. EV71 vaccine, a new tool to control outbreaks of hand, foot and mouth disease(HFMD)[J]. Expert Rev Vaccines, 2016, 15(5):599-606. DOI:10. 1586/14760584. 2016. 1138862.
[29]Wu JT, Jit M, Zheng Y, et al. Routine pediatric enterovirus 71 vaccination in China:a cost-effectiveness analysis[J]. PLoS Med, 2016, 13(3):e1001975.DOI:10. 1371/journal. pmed. 1001975.
[30]代敏，赵振希，张筱碟，等. 2010—2024年昆明市手足口病流行病学特征与病原学监测结果分析[J].热带病与寄生虫学，2025, 23(2):86-90, 115. DOI:10. 20199/j. issn. 1672-2302. 2025. 02. 005.
[31]Jiang L, Jiang H, Tian X, et al. Epidemiological characteristics of hand, foot, and mouth disease in Yunnan Province, China, 2008-2019[J]. BMC Infect Dis, 2021, 21(1):751. DOI:10. 1186/s12879-021-06462-4.
[32]Liu F, Yi Y, Song Y, et al. Epidemiology of hand,foot, and mouth disease outbreaks before and during availability of EV-A71 vaccine in China’s mainland:analysis of outbreak surveillance data from 2011 to 2023[J]. Lancet Reg Health West Pac, 2025, 59:101603.DOI:10. 1016/j. lanwpc. 2025. 101603.
[33]王泽宇，刘莹莹，赵文娜，等. EV-A71疫苗上市前后河北地区EV-A71流行株VP1区基因特征分析[J].病毒学报，2025,41(02):310-316. DOI:10. 13242/j. cnki.bingduxuebao. 250001.
[34]颜羽西，谢春燕，曹嘉钿，等.广东省EV-A71 B5基因型的分离鉴定、分子流行病学及抗原性研究[J].病毒学报，2025,41(06):1723-1730. DOI:10. 13242/j. cnki.bingduxuebao. 250218.
基本信息:
DOI：10.13242/j.cnki.bingduxuebao.250379
中图分类号:R725.1;R446.5
引用信息:
[1]刘泫果尔,姜黎黎,寸建萍,等.云南省昆明市2022-2023年手足口病病原谱及常见病原基因特征分析[J].病毒学报,2026,42(03):740-749.DOI:10.13242/j.cnki.bingduxuebao.250379.
2026-04-14
2026-04-14
2026-04-14