中国畜牧兽医2012，Vol.39Issue(8)：48-52,5.
细小病毒、猪伪狂犬病病毒和猪圆环病毒2型多重PCR检测方法的建立与应用
Establishment and Application of Multiplex PCR Assay for Detection of Porcine Parvovirus, Pseudorabies Virus and Porcine Circovirus Type 2
摘要
Abstract
Porcine parvovirus(PPV), pseudorabies virus(PRV) and porcine circovirus type 2CPCV2) were three main viral pathogens of porcine reproduction barrier diseases. To identify and differentiate rapidly the cause(s) of clinical diseases, a multiple PCR/RT-PCR assay was developed. Based on the similarity of the viral sequences deposited in the GenBank database, the VP2 gene of PPV, the gD gene of PRV and the ORF2 gene of PCV2 were selected as the diagnostic targets. By using three pairs of virus-specific primers, three PCR/RT-PCR assay were established to amplify the conservative regions of the three viruses, respectively. Consequently, a multiplex PCR/RT-PCR method to detect the three viruses in one tube was developed. The multiple PCR/RT-PCR system would amplify a 313 bp fragment for PPV, a 217 bp for PRV and a 447 bp for PCV2 simultaneously or separately in the samples, depending on its infection status. By comparison, the test proved that the multiplex PCR method being 100% coincidence with the single PCR, and it could be used for this three virus detection and differential diagnosis. 211 samples of pigs collected from 10 nosopoietic pig farms and outpatient cases were detected with the multiplex PCR assay established, the result showed that the positive rate of 42 positive PPV was 19. 91%, the positive rate of 26 positive PRV was 12. 32% and the positive rate of 56 positive PCV2 was 26. 54% , more than, the mixed infection rate of 25 two kinds of mixed infection was 11. 85%. It proved that the pigs had been infected by these three epidemics in Shanxi.关键词
猪细小病毒/猪伪狂犬病病毒/猪圆环病毒2型/多重PCR/建立与应用Key words
porcine parvovirus/ porcine pseudorabies virus/ porcine circovirus type 2/ multiplex PCR/ establishment and application分类
生物科学引用本文复制引用
王娟萍,姚敬明,赵喜有,吴忻,孟帆,刘文俊,樊振华,米瑞娟..细小病毒、猪伪狂犬病病毒和猪圆环病毒2型多重PCR检测方法的建立与应用[J].中国畜牧兽医,2012,39(8):48-52,5.基金项目
山西省科技厅攻关项目(20110311034-3). （20110311034-3）
