摘要
根据猪繁殖与呼吸综合征病毒（PRRSV）和猪瘟病毒（CSFV）的基因组序列设计了2对特异性引物,建立了同时检测PRRSV和CSFV的多重RT-PCR方法。2008年1月至2009年12月期间,采集山东各地71个不同规模猪场及农村散养户212份病料,采用建立的多重RT-PCR技术同时检测PRRSV和CSFV的感染状况。检测结果显示,病料中高致病性PRRSV占57.1%（121/212）,CSFV占29.2%（62/212）。应用RT-PCR方法扩增山东不同地区16株PRRSV的ORF5基因,并进行了序列分析。结果表明,16株PRRSV的ORF5基因核苷酸同源性为97.0%~100.0%;与参考变异毒株JXA1、传统毒株VR2332核苷酸同源性分别为98.0%~99.5%和86.4%~87.7%。推导的氨基酸序列分析表明,16株毒株GP5蛋白氨基酸不存在缺失,但存在位点突变。12株在非中和表位29位点与准种进化34位点发生了突变,分别由V→A和N→S;1株在非中和表位185位点发生突变,由A→V;这与2007年流行的PRRSV毒株JXA1、HUN等不同。
Two pairs of primers were designed according to the genome of CSFV and PRRSV,respectively,and a multiplex polymerase chain reaction（M-PCR） was established for the detection of the two viruses at the same time.A total of 212 samples were collected from 14 areas of shandong province during 2008-2009 and analysed using M-PCR.The result showed that the positive rates of PRRSV and CSFV were 57.1% and 29.2 %,respectively from Jan.2008 to Dec.2009.A total of 16 PRRSV strains were isolated from those samples and ORF5 gene was amplified and sequenced.The sequence analysis of ORF5 gene showed that the homology of ORF5 gene was up to 97.0%-100.0% among those 16 PRRSV isolates,and shared 98.0%-99.5% with JXA1,86.4%-87.7% with VR2332.A few of mutations but no deletions were found in deduced amino acid（aa） of ORF5 gene of 16 PRRSV isolates.Some key sites associated with antigenic epitopes,including 29th aa,34th aa and 185th aa have showed mutations,differing from JXA1,HUN,and other dominant isolates in 2007.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2011年第5期633-637,共5页
Chinese Journal of Veterinary Science
基金
山东省自然科学基金重点资助项目(Z2007D06)
山东省重大科技专项资助项目(2007ZHZX11103)
山东省农科院青年基金资助项目(2005YQ039)
