牛狂犬病病毒的鉴定及N基因序列分析

Identification and Sequence Analysis of Rabies Virus N Gene from Cattle

为诊断疑似牛狂犬病病例、分析其病原分子特征,本研究以疑似牛狂犬病脑组织为研究对象,运用内基氏小体染色观察、荧光抗体试验、特异性目的基因(N基因)RT-PCR扩增和序列测定等方法进行病毒鉴定,使用DNAStar、Mega4.0软件分析N基因的遗传进化关系。内基氏小体检测、荧光抗体试验、乳鼠脑内接种试验和特异性目的基因扩增结果显示狂犬病毒阳性,建立了基于狂犬病毒N基因的遗传进化关系树。结合临床症状确诊该病例为牛狂犬病,遗传进化分析表明本研究鉴定的狂犬病毒和2006年及2007年国内狗源狂犬病毒(DQ666306、DQ866121)、猪源狂犬病毒(DQ496219)及人源狂犬病毒(EF556197)的遗传关系较近。本文为研究牛狂犬病毒分子流行病学积累了资料。

To diagnose suspected cattle rabies case and analyze molecular characteristics of the pathogen, brain tissue materials of cattle with clinical symptom from a farm in China were used. Rabies virus （RV） in brain tissue was confirmed by Mouse Inoculation Test （MIT）, Hematoxy- lin and Eosin Staining （HE）, Fluorescent antibody test （FAT） and specific nucleoprotein gene （N） RT-PCR amplification. Molecular characterization of RV strain was performed using DNAS- tar, Mega4.0 softwares based on N gene sequence. Results showed that this cattle disease case was caused by RV, phylogenetic tree analysis demonstrated that RV researched in study has a closer genetic relationship with DQ666306, DQ866121, DQ496219, EF556197 which were isola- ted in 2006 and 2007 from dog, pig and human. This paper accumulated useful materials for fur- ther studying molecular epidemiology of cattle RV in China in the future.