检测猪圆环病毒2型DNA的套式PCR方法的建立及应用

为了进一步调查研究PVC2的流行病学及其致病机制,根据猪圆环病毒2型GD株及已发表的PCV2的全基因组序列,对来自广东、广西、湖南、海南等地287个猪场送检的1 560个可疑病料进行PCR扩增,并对扩增产物进行克隆、酶切、测序鉴定,建立了PCV2检测的套式PCR方法.结果表明,PCR扩增获得了预期大小的片断,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.该PCR方法对PCV2是特异的,并具有良好的重复性和稳定性.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,表明该病在我国广泛流行.

Establishment and Application of Nested PCR Assay for Testing PCV2

According to the published genome sequence of porcine circovirus type 1 and type 2(PCV1,.PCV2),1 560 doubtful diseased samples from 287 pig farms in Guangdong,Guangxi,Hunan,Hainan etc were made for PCR amplification.The amplified product was made for clone,enzyme restrictions test sequencing analysis and identification,then the nested polymerase chain reaction(nPCR) assay was established for PCV2.Results showed that PCR amplification got the factions that their size fixed in with expectation.Comparison of sequencing results and PCV2 sequence embodied by GenBank showed that the homology was more than 90%.The PCR assay was specificity to PCV2 and had good repeat and stability.1 560 samples from 287 pig farms had been detected for PCV2,from which 983 samples got the positive results for PCV2,suggesting that PCV2 infection is very popular in China.