河南省濮阳市手足口病病原检测及EV71基因序列特征分析  

Etiologic detection on HFMD and analysis of gene sequence features of EV71 in Puyang of Henan province

作　　者：徐国锋[1] 张清杰[1] 张玉华[1] 王爱梅[1] 孟晓飞[1] 卫海燕[2] 

机构地区：[1]濮阳市疾病预防控制中心,河南濮阳457000 [2]河南省疾病预防控制中心,河南郑州450016

出　　处：《中国卫生检验杂志》2015年第14期2377-2379,2382,共4页Chinese Journal of Health Laboratory Technology

基　　金：濮阳市科技局项目(130642)

摘　　要：目的对2010年-2012年濮阳市手足口病病例进行病原检测及EV71基因测序,明确致病病原。方法用real-time RT-PCR法对病例的咽拭子、肛拭子进行EV、EV71、Cox A16核酸检测,用RD细胞进行病毒分离,病毒培养物的PCR产物进行双向测序,用Mega 4.0生物软件完成全基因序列进化树构建。结果 2 097例手足口病,男女比例为1.90∶1,0岁~3岁儿童占90.61%。重症患者EV71阳性率为74.95%;普通患者EV71阳性率为23.14%,检测结果差异有统计学意义（χ2=562.144,P〈0.001）。2012年15份肛拭子EV71病毒分离阳性率为46.67%,15份咽拭子未分离出病毒。选取2012年1株重症患者EV71全基因序列构建系统进化树,显示与山东2010年莱阳株、阜阳2008年流行株聚成一簇同属C4a亚型。结论学龄前儿童是手足口病防控的重点,EV71病毒分离标本肛拭子优于咽拭子,濮阳市手足口病重症患者以EV71感染为主,为C4a亚型。Objective To conduct the etiologic detection on the patients with hand- foot- mouth disease（ HFMD） and the gene sequence features of EV71 in Puyang,Henan province from year 2010 to 2012,so as to identify the pathogen. Methods The throat swabs and rectal swabs of cases were detected for EV,EV71 and Cox A16 nucleic acids by real- time RT- PCR. Virus was isolated by using RD cells. PCR products of virus culture were undergoing bidirectional sequencing. The gene sequence phylogenetic tree was constructed by using Mega 4. 0 biological software. Results Among 2 097 patients of HFMD,male to female ratio was 1. 90∶1,and the numbers of 0- 3 years old children accounted for 90. 61%. The EV71 positive rate of severe patients was 74. 95%,and EV71 positive rate of ordinary patients was 23. 14%,the difference of detection results was statistically significant（ χ2= 562. 144,P〈0. 001）. The EV71 virus isolation positive rate of 15 patients of anal swabs was 46. 67% in 2012,but virus was not isolated in 15 patients of throat swabs. A entire gene sequence of severe EV71 patients was selected to build phylogenetic tree in 2012,and the result showed that it was the same with Shandong Laiyang strain in 2010 and Fuyang epidemic strain in 2008,which were belonged to subtype C4 a. Conclusion Preschoolers were HFMD prevention focus. The anal swab used to isolate EV71 virus was better than throat swab. The infection of severe cases of HFMD mainly was EV71,which belonged to C4 a subtype.

关 键 词：手足口病 核酸检测 肠道病毒71型 全基因序列 系统进化 

分 类 号：R512.5[医药卫生—内科学]