Isolation and identification of enterovirus 71 in endotracheal
aspirates from a patient with hand-foot-mouth disease

山东大学学报(医学版) ›› 2012, Vol. 50 ›› Issue (2): 106-109.

1. 山东省医学科学院基础医学研究所山东省罕少见病重点实验室，济南 250062；
2. 济南市传染病医院传染科，济南 250021

收稿日期:2011-11-04 出版日期:2012-02-10 发布日期:2012-02-10
通讯作者: 孟红（1960- ），女，研究员，主要从事医学病毒学的研究。E-mail: menghyky@yahoo.com.cn
作者简介:岳盈盈（1978- ），女，硕士，助理研究员，主要从事医学病毒学的研究
基金资助:
国家自然科学基金资助项目（81000720）；山东省自然科学基金资助项目（ZR2010HQ039）

Isolation and identification of enterovirus 71 in endotracheal
aspirates from a patient with hand-foot-mouth disease

YUE Ying-ying1, ZHANG Ying2, LI Peng1, LI Zhi-hui1,
SONG Nan-nan1, JI Xuan1, GAI Zhong-tao2, MENG Hong1

1. Institute of Basic Medicine，Shandong Academy of Medical Sciences,
Key Laboratory of Rare and Uncommon Diseases, Jinan 250062, China;
2. Department of Infection, Jinan Infectious Diseases Hospital, Jinan 250021, China

Received:2011-11-04 Online:2012-02-10 Published:2012-02-10

摘要/Abstract

摘要：

目的分离鉴定重症手足口病患者气管插管吸出液标本的EV71病毒株。方法将采集的标本过滤除菌接种于MA104细胞，出现细胞病变效应（CPE）后提取培养上清液中的RNA，用国家标准检测引物进行RT-PCR鉴定；用特异性引物进行PCR获得VP1全序列，测序后以MEGA4软件与各血清型代表株VP1序列进行分析并构建分子进化树。结果接种标本3d后细胞出现CPE，经RTPCR鉴定为EV71，对其VP1全序列进行测定分析后，确定其为C4亚型，并绘制了分子进化树。结论手足口病患者下呼吸道分泌物EV71的分离鉴定，为手足口病肺损伤发病机制的研究提供了依据。

关键词: 肠道病毒A型，人；手足口病；病毒培养

Abstract:

Objective To isolate and identify enterovirus 71(EV71) in the endotracheal aspirate specimen from a patient with hand-foot-mouth disease(HFMD). Methods The endotracheal aspirate specimen was inoculated in MA104 cells. Total RNA was extracted after the cytopathic effect (CPE) was observed, and the national standard primer was used to detect EV71 by RT-PCR. The VP1 complete sequence was obtained by PCR and then sequenced. The evolutionary tree was structured by MEGA4. Results Cells showed CPE 3 days after inoculation. The virus was identified as EV71 by RT-PCR. After the VP1 complete sequence was analyzed, the virus was confirmed as C4 serotype, and the evolutionary tree was drew by MEGA4. Conclusions For the first time, EV71 is isolated from lower respiratory tract secretion from HFMD patients, providing a basis for the study of pathogenesis of lung tissue injury in HFMD patients.

Key words: Enterovirus A, human; Hand, foot and mouth disease; Viruses cultivation

中图分类号:

R373.2

岳盈盈1，张颖2，李鹏1，李志会1，宋楠楠1，纪璇1，盖中涛2，孟红1. [J]. 山东大学学报(医学版), 2012, 50(2): 106-109.

YUE Ying-ying1, ZHANG Ying2, LI Peng1, LI Zhi-hui1, SONG Nan-nan1, JI Xuan1, GAI Zhong-tao2, MENG Hong1. Isolation and identification of enterovirus 71 in endotracheal
aspirates from a patient with hand-foot-mouth disease[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2012, 50(2): 106-109.

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Isolation and identification of enterovirus 71 in endotracheal
aspirates from a patient with hand-foot-mouth disease