1起由CV-A6型肠道病毒引起的手足口病疫情的快速鉴定与分析

doi:10.13217/j.scjpm.2015.0016

华南预防医学 ›› 2015, Vol. 41 ›› Issue (1): 16-20.doi: 10.13217/j.scjpm.2015.0016

1起由CV-A6型肠道病毒引起的手足口病疫情的快速鉴定与分析

曾汉日^1，2，李晖¹，郑焕英¹，刘冷¹，郭雪¹，管大伟¹，孙立梅¹，谭小华¹，柯昌文¹

1.广东省疾病预防控制中心，广东广州 511430; 2.中山大学公共卫生学院

修回日期:2015-03-30 出版日期:2015-02-20 发布日期:2015-03-30
作者简介:曾汉日（1980—），男，大学本科，主管技师，主要从事病原微生物检测工作

Rapid detection of an outbreak of hand, foot, and mouth disease caused by CV-A6 enterovirus

ZENG Han-ri^1,2, LI Hui¹, ZHENG Huan-ying¹, LIU Leng¹, GUO Xue¹, GUAN Da-wei¹, SUN Li-mei¹, TAN Xiao-hua¹, KE Chang-wen¹

1.Guangdong Provincial Center for Disease Control and Prevention, Guangzhou 511430, China; 2.School of Public Health, Sun Yat-Sen University

Revised:2015-03-30 Online:2015-02-20 Published:2015-03-30

摘要/Abstract

摘要： 目的运用分子生物学方法,对2013年5月河源市1所幼儿园发生的1起由CV-A6型肠道病毒引起的手足口病聚集性疫情进行肠道病毒型别快速鉴定。方法采集该起手足口病聚集性疫情的病例及密切接触者的粪便或咽拭子标本,运用荧光定量PCR方法进行总肠道病毒、EV-A71、CV-A16和CV-A6检测,并结合半巢式PCR方法进行肠道病毒VP1序列扩增,通过对目的条带序列测定及分析进行肠道病毒分型鉴定。结果经荧光定量PCR检测,26例病例35份标本中共有25例34份标本检出总肠道病毒阳性,38名密切接触者38份标本中有11人11份标本检出阳性,肠道病毒阳性率分别为96.15%和28.95%,其中分别有25例病例和7名密切接触者CV-A6阳性,阳性率分别为96.15%和18.42%,全部标本EV-A71和CV-A16检测阴性。7例病例和3名密切接触者的10份标本半巢式PCR扩增阳性,测序结果显示VP1序列均属于CV-A6,且核苷酸同源性为100.0%。结论广东省河源市此次发生于幼儿园的手足口病疫情由肠道病毒CV-A6型引起;荧光定量PCR与半巢式PCR扩增肠道病毒VP1序列相结合的方法有利于对引起手足口病的CV-A6型肠道病毒进行快速及准确鉴定。

Abstract: Objective To rapidly detect and identify an outbreak of hand, foot, and mouth disease (HFMD) caused by CV-A6 enterovirus in Heyuan City in May 2013 with molecular biological technology. Methods Specimens of stool and throat swabs of HFMD cases and close contacts were collected and detected for pan-EV, EV-A71, CV-A16, and CV-A6 by real-time PCR. VP1 genetic partial fragments of the positive samples were amplified with semi-nested PCR, and were adopted for sequencing and genetic analysis. Results In this outbreak, 96.15% of cases (25/26) and 28.95% of close contacts (11/38) were pan-EV positive; 96.15% of cases (25/26) and 18.42% of close contacts (7/38) were CV-A6 positive. All samples were negative for EV-A71 and CV-A16 nucleic acid. Partial fragments of VP1 were amplified from 10 samples of 7 cases and 3 close contacts by using semi-nested PCR. Sequence analysis showed that the nucleotide homology of all partial VP1 was 100.0% homologous with CV-A6. Conclusion The outbreak in a kindergarten in Heyuan City of Guangdong Province was caused by enterovirus CV-A6. Combining the real-time PCR and semi-nested PCR can be an effective way for rapid detection and identification of CV-A6 enterovirus.

中图分类号:

R183.4

曾汉日，李晖，郑焕英，刘冷，郭雪，管大伟，孙立梅，谭小华，柯昌文. 1起由CV-A6型肠道病毒引起的手足口病疫情的快速鉴定与分析[J]. 华南预防医学, 2015, 41(1): 16-20.

ZENG Han-ri, LI Hui, ZHENG Huan-ying, LIU Leng, GUO Xue, GUAN Da-wei, SUN Li-mei, TAN Xiao-hua, KE Chang-wen. Rapid detection of an outbreak of hand, foot, and mouth disease caused by CV-A6 enterovirus[J]. S China J Prev Med, 2015, 41(1): 16-20.

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1起由CV-A6型肠道病毒引起的手足口病疫情的快速鉴定与分析

Rapid detection of an outbreak of hand, foot, and mouth disease caused by CV-A6 enterovirus

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