猪传染性胃肠炎病毒的分离与鉴定

摘要

将经胶体金试纸条和RT-PCR鉴定为猪传染性胃肠炎病毒(TGEV)阳性的内蒙古某猪场的粪便样品,经ST细胞分离培养后,得到一株能产生明显细胞病变的毒株.纯净性检测结果显示,该毒株无细菌生长,无支原体及无外源病毒污染.特异性检测结果表明:该分离株能被TGEV特异性阳性血清中和,且能被TGEV FA荧光抗体识别.采用TGEV N基因特异性引物,经RT-PCR可扩增出1215 bp特异性片段,将该片段测序后与GenBank中已发表的13株TGEV N基因的序列进行同源性比较,同源性为98.1%~100%,其中与我国分离的H16、JS2012以及Miller M6毒株同源性关系最近,均为100%.结果表明,分离到的毒株为TGEV,命名为TGEV NMG株.%The feces of diarrheal piglets was detected by GIA and RT-PCR, the result showed that the sample was infected with TGEV. A strain of TGEV was isolated from ST cell, obvious cytopathic effect developed in the cell monolayer. The purity test indicated that the virus was free from the bacteria, mycoplasma and extra viruses infection. The specification test manifested that the strain could be neutralized by anti-TGEV serum and recognized by direct immunofluorescence antibody assay. The target fragment(1215 bp)was amplified by RT-PCR and sequenced,and its sequence was compared with other 13 TGEV strains in GenBank database. The result showed that the nucleotide homology is in 98.1%~100%, and the NMG strain showed 100% identity to starins H16、JS2012 and Miller M6. These results showed that a strain of TGEV was isolated,and named TGEN NMG.