牛病毒性腹泻病毒增殖特性研究

Study on the Proliferation of Bovine Viral Diarrhea Virus

为研究牛的病毒性腹泻病毒(BVDV)的增殖特性,从南充地区腹泻牛的胃内容物提取病毒样本,利用MDBK细胞分离出毒株,通过RT-PCR进行扩繁检测、基因序列对比和病毒试验,最终确定分离菌株为腹泻病毒。选择BVDV病毒感染MDBK细胞,设定为感染的对照组,在感染48 h后,收集细胞的病毒核酸,采用实时荧光定量PCR进行病毒感染RNA进行定量分析,确定MDBK细胞感染后的细胞病毒生长曲线。结果表明,感染后2~4 h,病毒的RNA增殖较慢,4~24 h出现倍增,24~72 h复制变缓,保持在一定水平。这说明BVDV感染MDBK细胞后,病毒在细胞内出现增殖,并随着感染时间的推移,病毒数量趋于稳定。表明BVDV感染细胞,病毒在细胞内增殖后会向细胞外释放。

In order to study the proliferative characteristics of bovine viral diarrhea virus(BVDV),a strain of BVDV was isolated from the stomach contents of diarrheal cattle in Nanchong,the isolated strain was confirmed to be diarrheal virus by RTPCR detection and amplification,gene sequence comparison and virus test.The MDBK cells were infected with BVDV virus.The viral nucleic acid of the cells was collected 48 hours after infection and the viral RNA was Quantitative analysis by real-time PCR,to determine the virus growth curve of MDBK cells after infection.The results showed that the viral RNA proliferated slowly from 2 to 4 hours after infection,doubled from 4 to 24 hours,and slowed down from 24 to 72 hours after infection,and remained at a certain level.This indicated that the virus proliferated in MDBK cells after being infected by BVDV,and the number of virus tended to be stable with the passage of time. The results showed that BVDV could infect the cells,and the virus could be released into the cells after proliferating.