PCV2a与PCV2b鉴别诊断PCR方法的优化及应用  

OPTOMIZATION AND APPLICATION OF A DIFFERENTIAL PCR ASSAY FOR DETECTION OF PORCINE CIRCOVIRUS TYPE 2A AND 2B

作　　者：翟少伦[1,2] 张建武[1] 韦祖樟[1] 袁世山[1] 岳城[2] 冉多良[2] 龙进学[1] 

机构地区：[1]中国农业科学院上海兽医研究所,上海200241 [2]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《中国动物传染病学报》2010年第1期1-7,共7页Chinese Journal of Animal Infectious Diseases

基　　金：十一五支撑项目(2006BAD06A04)

摘　　要：猪圆环病毒2型(Porcine circovirus type 2,PCV2)是严重危害养猪业的重要病原之一,根据基因组大小可分为2个基因型:PCV2a和PCV2b。为更好地掌握PCV2a和PCV2b在我国猪场的流行情况,本研究对先前的鉴别PCR诊断方法(涉及到引物、PCR体系及程序)进行优化,并应用优化的鉴别PCR方法对118份PCV2阳性样品进行检测,结果显示PCV2a占30.5%(36/118),PCV2b占37.3%(44/118),共感染占32.2%(38/118),与常规PCR检测结果符合率为100%,此外,对30份临床送检组织和血清样品也有很高的检出率。部分鉴别PCR扩增的目的产物进行测序及序列进化树分析,表明扩增序列为PCV2a与PCV2b的特征序列。经本研究优化后的鉴别PCR方法易操作、特异性强、敏感性好,可广泛用于临床样品中PCV2a和PCV2b的快速检测。Abstract： Porcine circovirus type 2 （PCV2） is one of the most important pathogenic agents for swine industry, which is divided into two genotypes （PCV2a and PCV2b） according to their genome sizes. To accurately investigate into molecular epidemiology of PCV2a and PCV2b in Chinese swine herds, a differential PCR assay （D-PCR） that was previously described was optimized in the present study for primers, reaction system and program. The optimized D-PCR was used to test 118 PCV2 positive samples. Results showed that single infection rates of either PCV2a or PCV2b and co-infection of PCV2a/PCV2b were 30.5 % （36/118）, 37.3 % （44/118） and 32.2 % （38/118）, respectively. These results were in agreement with ordinary PCR detection. High percentage of PCV2a or PCV2b infection was also found in 30 clinical tissue and serum samples as tested by D-PCR. In addition, the products of D-PCR showed characteristic sequences of PCV2a and PCV2b by sequencing and phylogenetic tree analysis. Optimized D-PCR is easily operational, highly specific and sensitive, which can be applied for rapid differentiation of PCV2a and PCV2b in clinical samples.

关 键 词：猪圆环病毒2型 鉴别诊断 PCR 

分 类 号：S858.282.659.2[农业科学—临床兽医学] Q7[农业科学—兽医学]