云南省昆明地区2022年柯萨奇病毒A16型的VP1区分子特征

张磊; 沈茹; 楚昭阳; 马绍辉; 李丽

doi:10.12259/j.issn.2095-610X.S20240711

云南省昆明地区2022年柯萨奇病毒A16型的VP1区分子特征

doi: 10.12259/j.issn.2095-610X.S20240711

张磊¹,
沈茹¹,
楚昭阳²,
马绍辉²,
李丽^1, ,

1.
昆明市妇幼保健院检验科，云南昆明　650031
2.
中国医学科学院/北京协和医学院医学生物学研究所/云南省重大传染病疫苗研发重点实验室，云南昆明　650118

基金项目: 云南省科技计划项目（202202AA100016）；昆明市卫生健康委科研项目（2022-11-01-001）；科技人才与平台计划（ 202005AD160025）

详细信息

作者简介:
张磊（1989～），女，云南昆明人，医学学士，主管检验师，主要从事生物化学检测工作

通讯作者:
李丽，E-mail：zhuoyuell@163.com

中图分类号: 373.2
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- 被引次数: 0
出版历程
- 收稿日期: 2024-01-22
- 网络出版日期: 2024-06-14
- 刊出日期: 2024-07-25

Molecular Characterization of the VP1 Region of Coxsackievirus 16 in Kunming，Yunnan in 2022

1.
Dept. of Clinical Laboratory，Kunming Maternal and Child Health hospital，Kunming Yunnan 650031
2.
Institute of Medical Biology，Chinese Academy of Medical Sciences and Peking Union Medical College，Key Laboratory for Research and Development of Major Infectious Diseases Vaccine in Yunnan，Kunming Yunnan 650118，China

摘要

摘要: 目的对昆明市某医院2022年从手足口病患儿分离的柯萨奇病毒A组16型（coxsackievirus A16，CVA16）的全VP1基因区分子特征分析。方法对昆明市某医院2022年从手足口病（hand foot mouth disease，HFMD）患儿200份临床样品粪便标本进行处理，分别通过RD细胞和Vero细胞分离，对经3次培养后仍产生细胞效应（cytopathogenic effect，CPE）的细胞培养液提取病毒核酸，然后用通用引物222/224经RT-PCR扩增并测序，序列经BLAST比对确定其血清型，对鉴定为柯萨奇病毒A组16型（coxsackievirus A16，CVA16）的再用特异引物CA16VP1F/CA16VP1R扩增其全VP1区并测序和拼接。下载CVA16病毒标准序列，采用Geneious 5.4.1和MEGA 6.05等软件对本研究CVA16的全VP1区的核苷酸和氨基酸序列分析。结果经鉴定后共检出22株从Vero细胞分离CVA16和6株RD细胞分离的CVA16毒株，进一步通过特异引物扩增获得21株可用的CVA16 VP1全长序列，基于全VP1基因系统进化分析，这21株CVA16全属于B1a基因型。21株CVA16全长VP1基因之间的核苷酸和氨基酸序列同源性均较高，分别为91.9%～99.6%和89.2%～99.3%；与中国CVA16 B1a基因型的其他株的核苷酸和氨基酸序列同源性分别为89.7%～96.3%和89.2%～97.0%。与CVA16 B1b其他参考株对比，其核苷酸和氨基酸序列同源性分别在84.3%～87.2% 和89.2%～97.0%。与其它CVA16的B1a基因亚型相比较发现21个氨基酸位点出现变异。结论 2022年中国云南省昆明某医院引起HFMD的CVA16毒株属于B1a基因型，应继续监测以明确其流行特征。
- 柯萨奇病毒A组16型 /
- 全VP1基因 /
- 序列分析
Abstract: Objective To analyze the molecular characteristics of the whole VP1 gene region of coxsackievirus A16 （CVA16） isolated from children with hand, foot and mouth disease （HFMD） in 2022 from a hospital in Kunming, Yunnan. Methods Two hundreds fecal specimens of HFMD clinical samples from 2022 in a hospital in kunming, were processed and isolated by RD cells and Vero cells, respectively, and viral nucleic acids were extracted from the cell cultures that still produced cytopathogenic effect （CPE） after three times of incubation, then amplified and sequenced by RT- PCR using universal primers 222/224, and sequenced, and the sequences were compared by BLAST to determine the serotypes, and then the full VP1 region of CVA16 was amplified with specific primers CA16VP1F/CA16VP1R , then sequenced and spliced. The standard sequences of CVA16 virus were downloaded, and the nucleotide and amino acid sequences of the whole VP1 region of CVA16 in this study were analyzed using software such as Geneious 5.4.1 and Mega 6.05. Results 22 strains of CVA16 isolated from Vero cells and 6 strains isolated from RD cells were detected by RT-PCR using universal primers, and the available sequences of 21 strains of full-length CVA16 VP1 region were further amplified by the specific primers of CVA16, and the nucleotide and amino acid sequences of the 21 strains of full-length CVA16 VP1 genes were highly homologous to each other, with 91.9%～99.6% and 89.2%～99.3%, respectively. The nucleotide and amino acid sequence homology with other strains of the chinese B1a genotype ranged from 89.7%～96.3% and 94.6%～99.0%, respectively; and nucleotide and amino acid sequence homology was in the range of 84.3%～87.2% and 89.2%～97.0%, respectively, when compared with the other reference strains of B1b. The phylogenetic analysis showed that all 21 CVA16 strains belonged to the B1a genotype. The 21 mutations were found by compared with that of other B1a strains. Conclusions The CVA16 strain that caused HFMD in a hospital in Kunming, Yunnan Province, China in 2022 belongs to the B1a genotype. Surveillance should be continued to clarify its epidemiological characteristics.
- Coxsackievirus A16 /
- Complete VP1 gene /
- Sequence analysis

HTML全文

图 1 CVA16全VP1区推测氨基酸序列的比对

Figure 1. Comparison of putative amino acid sequences of the full VP1 region of CVA16 strains

下载: 全尺寸图片幻灯片

图 2 基于全VP1区核苷酸序列的CVA16的系统进化树

Figure 2. Phylogenetic tree based on full VP1 sequences of CVA16 strains

下载: 全尺寸图片幻灯片

参考文献(25)

[1]	Liu Y,Chen J,Zhang M,et al. Coxsackievirus B: the important agent of hand,foot,and mouth disease[J]. Journal of Medical Virology,2023,95(3):e28669. doi: 10.1002/jmv.28669
[2]	Fu X,Wan Z,Li Y,et al. National epidemiology and evolutionary history of four hand,foot and mouth disease-related enteroviruses in China from 2008 to 2016[J]. Virologica Sinica,2020,35(1):21-33. doi: 10.1007/s12250-019-00169-2
[3]	冯骁,李琦涵. 柯萨奇病毒A16型疫苗的研究进展[J]. 中国生物制品学杂志,2021,34(3):357-361.
[4]	Noisumdaeng P,Korkusol A,Prasertsopon J,et al. Longitudinal study on enterovirus A71 and coxsackievirus A16 genotype/subgenotype replacements in hand,foot and mouth disease patients in Thailand,2000–2017[J]. International Journal of Infectious Diseases,2019,80(1):84-91.
[5]	Hoa-Tran T N,Dao A T H,Nguyen A T,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
[6]	Ji H,Fan H,Lu P,et al. Surveillance for severe hand,foot,and mouth disease from 2009 to 2015 in Jiangsu province: epidemiology,etiology,and disease burden[J]. BMC Infectious Diseases,2019,19(1):1-18. doi: 10.1186/s12879-018-3567-x
[7]	Cai K,Wang Y,Guo Z,et al. Clinical characteristics and managements of severe hand,foot and mouth disease caused by enterovirus A71 and coxsackievirus A16 in Shanghai,China[J]. BMC infectious diseases,2019,19(1):1-8. doi: 10.1186/s12879-019-3878-6
[8]	Zhang M,Zhao Y,Zhang H,et al. Molecular characterization of Coxsackievirus A16 strains isolated from children with severe hand,foot,and mouth disease in Yunnan,Southwest China,during 2009‐2015[J]. Journal of Medical Virology,2019,91(1):155-160. doi: 10.1002/jmv.25297
[9]	Chen L,Yao X J,Xu S J,et al. Molecular surveillance of coxsackievirus A16 reveals the emergence of a new clade in mainland China[J]. Archives of virology,2019,164(3):867-874. doi: 10.1007/s00705-018-4112-3
[10]	Han Z,Song Y,Xiao J,et al. Genomic epidemiology of coxsackievirus A16 in mainland of China,2000–18[J]. Virus evolution,2020,6(2):1-15.
[11]	Yi L,Zeng H,Zheng H,et al. Molecular surveillance of coxsackievirus A16 in southern China,2008–2019[J]. Archives of Virology,2021,166(6):1653-1659. doi: 10.1007/s00705-021-05052-8
[12]	Guo J,Cao Z,Liu H,et al. Epidemiology of hand,foot,and mouth disease and the genetic characteristics of Coxsackievirus A16 in Taiyuan,Shanxi,China from 2010 to 2021[J]. Frontiers in cellular and infection microbiology,2022,12(1):1040414.
[13]	Jiang L,Jiang H,Tian X,et al. Epidemiological characteristics of hand,foot,and mouth disease in Yunnan Province,China,2008–2019[J]. BMC infectious diseases,2021,21(1):1-8. doi: 10.1186/s12879-020-05706-z
[14]	KangY J,Shi C,Zhou J,et al. Multiple molecular characteristics of circulating enterovirus types among paediatric hand,foot and mouth disease patients after EV71 vaccination campaign in Wuxi,China[J]. Epidemiology & Infection,2022,150(1):1-19.
[15]	Jiang H,Zhang Z,Rao Q,et al. The epidemiological characteristics of enterovirus infection before and after the use of enterovirus 71 inactivated vaccine in Kunming,China[J]. Emerging Microbes & Infections,2021,10(1):619-628.
[16]	Wang M,Chen T,Peng J,et al. The spatial–temporal distribution and etiological characteristics of hand-foot-and-mouth disease before and after EV-A71 vaccination in Kunming,China,2017-2020[J]. Scientific Reports,2022,12(1):17028. doi: 10.1038/s41598-022-21312-2
[17]	Song C,Li Y,Zhou Y,et al. Enterovirus genomic load and disease severity among children hospitalised with hand,foot and mouth disease[J]. E Bio Medicine.,2020,62(1):103078.
[18]	杨溪,曹亿会,寸建萍,等. 2015—2019年云南省手足口病流行及病原学特征分析[J]. 职业卫生与病伤,2022,37(5):278-282.
[19]	姜法春,赵超,董鸿铭,等. 2019年冬季青岛市手足口病患者感染柯萨奇病毒A组16型基因特征分析[J]. 疾病监测,2022,37(6):755-759.
[20]	杜燕华,关路媛,冯瑄,等. 2019年陕西省手足口病的病原构成及柯萨奇A16型病毒的VP1区序列分析[J]. 现代预防医学,2022,49(7):1289-1293.
[21]	嵇红,鲍倡俊,祁贤,等. 2009-2017年江苏省手足口病病原谱及柯萨奇A16型的分子流行病学研究[J]. 中国人兽共患病学报,2019,35(2):113-119.
[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]	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):1026-1031.
[24]	刘艳艳,姜黎黎,李文龙,等. 2021年昆明市手足口病流行病学和肠道病毒病原谱特征[J/OL]. 中国疫苗和免疫,2023,29(1): 64-70.
[25]	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型基因特征分析[J]. 中国热带医学,2023,23(10):1104-1108.

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