猪瘟病毒GZA株的分离鉴定及遗传变异分析

Isolation and identification of classical swine fever virus GZA strain and genetic variation

本研究收集贵州规模化猪场猪瘟净化过程中RT-PCR检测阳性病料,采用细胞接毒技术、病毒的形态结构观察和间接免疫荧光试验等分离鉴定了1株猪瘟病毒(CSFV),命名为CSFV GZA株,并对GZA株E2基因的核苷酸和氨基酸序列进行差异性分析。分离鉴定结果表明:接种PK-15细胞后连传7代RT-PCR均能扩增出CSFV特异性条带;病毒粒子在电镜下呈椭圆形或圆形,直径30～80nm;该毒株E2基因与参考毒株E2基因的核苷酸同源性82.8%～83.4%,氨基酸相似度88.7%～90.6%,并在713,725,729,734,738氨基酸位点发生改变;遗传进化树分析还得出GZA株E2基因与参考毒株遗传距离较远。说明本次分离株与其他流行株存在一定的差异,可为以后CSFV病原学的研究提供参考。

A classical swine fever virus was isolated from the positive sample of large scale pig farm in Guizhou and identified by RT-PCR and sequence analysis.The classical swine fever virus were observed in infected cell by electron microscope and the E2gene sequence was analyzed.The alignment of the E2sequence showed that the isolate shared high similarity(82.8%-83.4%)with other CSFV reference strains available in GenBank.These results could be useful for the research of epidemiology and molecular biology of CSFV.