摘要
为了解山东滨州某猪场PRRSV的流行及变异情况,将该猪场采集的疑似PRRSV阳性病料进行RT-PCR检测,检测出的阳性样品用Marc-145细胞进行分离,设计了9对扩增引物用于分离株(BZ4-19)的全基因扩增,将扩增出的全基因序列进行核苷酸及其推导的氨基酸序列分析。最终成功在Marc-145细胞上分离到1株RespPRRS MLV-like毒株,全基因扩增结果显示其全长为15412 bp(不含polyA尾);核苷酸同源性分析说明,其与美洲经典株VR2332及RespPRRS MLV(疫苗株)同源性最高,分别为99.4%和99.8%。将BZ4-19分离株与VR2332、RespPRRS MLV的氨基酸序列进行比较分析揭示,VR2332与RespPRRS MLV共有30处氨基酸差异,在这30处差异中,分离株与VR2332有7处一致,与RespPRRS MLV有23处一致。在鉴别VR2332与RespPRRS MLV的9处可能与毒力相关的氨基酸位点时,分离株有7处与VR2332相同,由此推测本研究分离到的毒株属于疫苗株RespPRRS MLV返强毒株,我国具有上述分子变化的PRRSV的流行应被关注。
To understand the prevalence and variation of PRRSV in a pig farm in Binzhou,Shandong Province,China,suspected PRRSV-positive material collected from this farm was subjected to RT-PCR.The detected positive samples were isolated using Marc-145 cells,then nine pairs of amplification primers were designed for whole gene amplification of the isolate(named BZ4-19),and the amplified whole gene sequence was analyzed for nucleotides and their deduced amino acid sequences.The results showed that a RespPRRS MLV-like strain was successfully isolated from Marc-145 cells,and the whole gene amplification showed that its full length was 15412 bp(without polyA).Nucleotide homology analysis showed that it had the highest homology with the American classical strain VR2332 and RespPRRS MLV(vaccine strain),with 99.4%and 99.8%.Amino acid sequences of BZ4-19 isolates were analyzed in comparison with those of VR2332,RespPRRS MLV,the results showed that VR2332 and RespPRRS MLV had a total of 30 amino acid differences.Of these 30 differences,the isolate was consistent with VR2332 in 7 instances and with RespPRRS MLV in 23 instances.In the identification of nine amino acid sites that may be related to virulence between VR2332 and RespPRRS MLV,seven of the isolates were identical to VR2332.It is hypothesized that the strain isolated in this study belongs to the vaccine strain RespPRRS MLV returned strong strain,and the prevalence of PRRSV with the above molecular changes in China should be concerned.
作者
李思琪
刘莹
刘濮毓
任晓祥
李岩岩
王文钊
左玉柱
范京惠
LI Siqi;LIU Ying;LIU Puyu;REN Xiaoxiang;LI Yanyan;WANG Wenzhao;ZUO Yuzhu;FAN Jinghui(College of Animal Medicine,Hebei Agricultural University,Baoding 071001,China;Hebei Veterinary Biotechnology Innovation Center,Baoding 071001,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2024年第2期200-207,共8页
Chinese Veterinary Science
基金
河北省农业产业技术体系生猪创新团队项目(HBCT2024220201,HBCT2024220401)
石家庄市科技计划项目(229500232N)。
