基础医学与临床 ›› 2019, Vol. 39 ›› Issue (3): 348-352.

• 研究论文 • 上一篇    下一篇

九江地区手足口病病原柯萨奇A6病毒的分离、鉴定及分子流行病学特征

余文敏1,柯秀梅1,邬亚华2,殷嫦嫦3,张义平1,朱振国4,徐焕新5   

  1. 1. 九江学院基础医学院
    2. 九江学院
    3. 九江学院/九江市转化医学重点实验室
    4. 南昌大学医学部
    5. 江西省九江市第三人民医院
  • 收稿日期:2018-04-28 修回日期:2018-07-17 出版日期:2019-03-05 发布日期:2019-03-05
  • 通讯作者: 余文敏 E-mail:yuwenmin2013@126.com
  • 基金资助:
    coxsackievirus A6 virus (CVA6); identification; epidemiological characteristics

Isolation, identification and molecular epidemiological characteristics of Coxsackie virus A6 in Jiujiang area

  • Received:2018-04-28 Revised:2018-07-17 Online:2019-03-05 Published:2019-03-05
  • Contact: min wenyu E-mail:yuwenmin2013@126.com

摘要: 目的 探讨九江地区流行手足口病病原体柯萨奇A6(CVA6)的分离、鉴定及分子流行特征。方法 取手足口病患儿的咽拭子,将咽拭子病毒悬液接种于人横纹肌肉瘤细胞(RD细胞)。通过CVA6病毒致细胞病变效应(CPE)分析、MTT法检测病毒增殖效果、CVA6病毒特异性核酸聚合酶链反应(PCR)及同源和遗传进化分析等方法检测CVA6的致病效果和分子流行特征。结果 咽拭子病毒悬液接种RD细胞盲传5代后出现CPE,收集其病毒培养基上清,再接种RD细胞2次,均出现CPE视为CPE阳性;CVA6病毒VP1区域通过序列比对,以此确认2株病毒为CVA6,分别命名JJ2012037和JJ2012039;CVA6呈时间依赖性抑制宿主细胞增殖;利用CVA6毒株的VP1序列和国内外流行的CVA6序列进行核苷酸同源性分析;该毒株与上海分离到CVA6(JX495144)序列发生的同源性最高(99.2%~99.6%)。结论 成功分离出2株九江地区流行的手足口病病原CVA6病毒株,这2株CVA6分离株的分子流行特征接近上海地区的CVA6的流行病原,可能为同一个基因亚型。

关键词: 柯萨奇A6病毒, 鉴定, 流行特征

Abstract: Objective To investigate the isolation,identification and molecular epidemiological characteristics of the CVA6 of hand foot mouth disease (HFMD) epidemic in Jiujiang.Methods Throat swabs from children with hand foot mouth disease were collected and the suspends was inoculated into RD cells.The isolation and pathogenicity of CVA6 were detected by CVA6 virus-induced cytopathic effect (CPE) analysis,MTT assay,CVA6 virus-specific nucleic acid polymerase chain reaction (PCR), and homology and phylogenetic analysis.Results The CPE cells were found after 5 generations in RD cells incubated with the supernatant. RD cells were infected with the supernatant collected for 2 consecutive infections, and the same CPE appeared. The VP1 region of the CVA6 virus was identified by sequence alignment to confirm that the viruses were CVA6, named JJ2012037 and JJ2012039 respectively.The MTT detection showed that CVA6 could inhibit the proliferation of host cells in a time-dependent manner. Nucleotide sequence homology analysis was carried out by using VP1 sequences of CVA6 strains from different countries and regions.The results showed that the homology of the CVA6 strain was as high as 99.2-99.6% with the strain separated from Shanghai.Conclusion Two strains of the epidemic CVA6 strain of hand-foot-mouth disease have been successfully isolated.The molecular epidemiology of the two CVA6 strains is close to the epidemic of CVA6 from shanghai, which may be the same subtype of the gene.

Key words: coxsackievirus A6 virus (CVA6), identification, epidemiological characteristics