1株高致病性血凝性脑脊髓炎病毒的分离与鉴定

Isolation and identification of a high pathogenicity hemagglutinating encephalom yelitis virus

利用细胞培养方法从临床表现神经症状的死亡仔猪脑组织内分离获得1株病毒,同时对该病毒进行电镜负染观察、昆明种小鼠致病性试验、RT-PCR检测和部分基因测序分析。电镜观察可见细胞培养物中有大量的直径约115 nm的冠状病毒样粒子;接种病毒的小鼠均出现典型的神经症状,死亡率100%;血凝性脑脊髓炎病毒(HEV)特异性引物RT-PCR扩增结果阳性;其S基因序列与GenBank中登录的HEV-67 N相应基因序列的同源性高达99.6%。结果表明,分离的病毒为猪血凝性脑脊髓炎病毒,分离株暂命名为HEV-CC-2007。

A high pathogenicity hemagglutinating encephalomyelitis virus （HEV） strain was successfully isolated from brain tissues of died pigs with clinical nerve symptom from a pig farm of Changchun in China by cells cul ture. The viruses were identified by electron microscope,Kunming mice regression test ,RT-PCR and partial gene sequences analysis. Electron microscope revealed the coronavirus-like virions that measured approximately 115 nm in diameter,all the mice were died after inoculating the viruses ,and the specific amplified fragments were obtained by RT-PCR of HEV primers. Subsequent analysis indicated the sequence of partial S gene of the virus shared a 99. 6% similarity with the standard strain of HEV-67N in GenBank. The results showed that the strain was a high pathogenicity HEV strain,and mainland of China,and was named HEV-CC-2007.