PCV2套式PCR检测方法的建立及应用  

Establishment and Application of Nested PCR Assay for PCV2 Detection

作　　者：靳玉芬[1] 郝贵增[1] 何宏轩[2] 

机构地区：[1]安阳工学院生物与食品工程学院,安阳455000 [2]中国科学院动物研究所,北京100091

出　　处：《吉林农业大学学报》2007年第4期437-439,446,共4页Journal of Jilin Agricultural University

基　　金：中国博士后科学基金(2003033120)

摘　　要：根据猪圆环病毒2型GD株及已发表的PCV2的全基因组序列,设计1对PCV2型特异性引物,对可疑病料进行PCR扩增。将扩增产物连接到pMD 18-T载体上并克隆到大肠杆菌DH5α中,提取质粒进行套式PCR、酶切、测序鉴定,结果扩增出了PCV2的目的片段。将测序结果与GenBank收录的PCV2序列进行比较,发现同源性均在90%以上。用该PCR方法对287个猪场的1 560份样品进行了检测,其中983份为PCV2阳性,阳性率为63%。According to the published genome of porcine circovirus type 1 and type 2（PCV1, PCV2）, two pairs of special primers were designed to detect PCV2 in samples from some farms by nested PCR assay. The PCR product was extracted and subcloned into pMD 18-T vector and then cloned into E. coli DI-ISa. The recombination plasmid was identified by digestion with different enzyme restrictions and sequencing analysis. The homology of nucleotide sequence between the PCR product and that of other PCV2 strains was more than 90%. Using this PCR method, 1 560 samples from 287 pig farms have been detected for PCV2, from which 983 samples got the positive results for PCV2, suggesting that PCV2 infection is very popular in China.

关 键 词：猪圆环病毒2型 套式PCR 克隆 PCR检测 

分 类 号：S852.65[农业科学—基础兽医学]