猪脑心肌炎病毒GXLC株的分离及其3D基因分子特征的分析

›› 2011, Vol. 38 ›› Issue (1): 81-87.

猪脑心肌炎病毒GXLC株的分离及其3D基因分子特征的分析

陈进喜1，施开创2，黄胜斌2，陆文俊2，屈素洁2，郑敏2，李军2

（1.广西钦州市动物疫病预防控制中心，广西钦州 535000； 2.广西动物疫病预防控制中心，广西南宁 530001）

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-01-01 发布日期:2011-01-01
通讯作者: 施开创

Isolation of Porcine Encephalomyocarditis Virus GXLC Strain and Analysis of the Molecular Characteristics of its 3D Gene

CHEN Jin-xi1，SHI Kai-chuang2，HUANG Sheng-bin2，LU Wen-jun2，QU Su-jie2，ZHENG Min2，LI Jun2

（1. Qinzhou Center for Animal Disease Control and Prevention，Qinzhou 535000，China;2. Guangxi Center for Animal Disease Control and Prevention，Nanning 530001，China）

Received:1900-01-01 Revised:1900-01-01 Online:2011-01-01 Published:2011-01-01
Contact: SHI Kai-chuang

摘要/Abstract

摘要： 采集临床疑似脑心肌炎死亡仔猪的组织作为接种材料，接种于BHK-21细胞系，观察细胞病变（CPE），并用RT-PCR和间接荧光抗体试验（IFA）进行鉴定，证实分离到1株脑心肌炎病毒（encephalomyocarditis virus，EMCV），命名为GXLC株。应用RT-PCR方法扩增GXLC株的3D基因，扩增产物克隆入pMD18-T载体后进行测序，对获得的3D基因序列进行分析。序列分析结果表明，GXLC株3D基因全长1380 nt，编码460个氨基酸，含有7个抗原表位。同源性分析结果表明，GXLC株与国内外其它EMCV分离株3D基因核苷酸序列的同源性在84.7%～99.7%之间，氨基酸序列的同源性在96.1%～99.6%之间。遗传进化分析结果表明，基于3D基因核苷酸序列绘制的系统进化树可将所有EMCV分离株分成2个群：Ⅰ群和Ⅱ群，Ⅰ群可再细分为Ⅰa亚群和Ⅰb亚群，其中猪源EMCV在Ⅰ群和Ⅱ群中均有分布，而鼠源EMCV分布在Ⅰ群，人源EMCV分布在Ⅱ群；GXLC株与其它中国分离株均属于Ⅰa亚群。

关键词: 猪; 脑心肌炎病毒; 分离; 3D基因; 分子特征

Abstract:

The clinical tissue samples collected from piglets died of suspected encephalomyocarditis were homogenated and the supernatant was inoculated into baby hamster kidney (BHK-21) cell lines to isolate encephalomyocarditis virus (EMCV). According to the cytopathic effect (CPE) and the results of RT-PCR and indirect fluorescent antibody test (IFA)，one strain of EMCV was successfully isolated and designated as GXLC strain. The 3D gene of GXLC strain was amplified by RT-PCR，cloned into pMD18-T vector，sequenced and analyzed. The sequence analysis showed that the 3D gene of GXLC strain consisted of 1380 nucleotides encoding 460 amino acids and contained 7 epitopes. The homology analysis indicated that the homology of 3D gene of GXLC strain and other EMCV strains available in GenBank were from 84.7% to 99.7% and from 96.1% to 99.6% based on the nucleotide and deduced amino acide sequences，respectively. The phylogenetic tree based on the 3D gene nucleotide sequences revealed that all the EMCV strains could be divided into two groups: group Ⅰ and group Ⅱ，and group Ⅰ could be subdivided into subgroup Ⅰa and subgroup Ⅰb. The strains from pig distributed in group I and group Ⅱ，while the strains from mouse distributed in group I and the strains from human distributed in group Ⅱ. GXLC strain，and other EMCV isolates from China belonged to subgroup Ⅰa.

Key words: swine; encephalomyocarditis virus; isolation; 3D gene; molecular characteristic

中图分类号:

陈进喜;施开创;黄胜斌;陆文俊;屈素洁;郑敏;李军. 猪脑心肌炎病毒GXLC株的分离及其3D基因分子特征的分析[J]. , 2011, 38(1): 81-87.

CHEN Jin-xi;SHI Kai-chuang;HUANG Sheng-bin;LU Wen-jun;QU Su-jie;ZHENG Min;LI Jun. Isolation of Porcine Encephalomyocarditis Virus GXLC Strain and Analysis of the Molecular Characteristics of its 3D Gene[J]. , 2011, 38(1): 81-87.

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