猪圆环病毒2型PCR检测方法的建立及其在临床检测中的应用

选择猪圆环病毒2型(PCV-2)基因保守区设计1对引物P1和P2,扩增536 bp的片段,该方法可以特异地检测出PCV-2的DNA,而对猪细小病毒、猪瘟病毒、猪伪狂犬病毒及未接PCV-2的PK-15细胞均呈阴性;该法能检测出29 ng/L的病毒DNA。对扩增片段进行测序,结果表明扩增片段属于PCV-2。应用该方法对2008年度上海及周边地区送检的91份临床样本进行了PCV-2的检测,结果表明,阳性样本为52份,阳性率为57.14%。研究结果表明,建立的PCR方法检测PCV-2具有较好的敏感性和特异性,可用于PCV-2感染疑似病例的诊断及其分子流行病学调查。

Development of PCR to Detect Porcine Circovirus Type 2 and its Clinical Application

As porcine circovirus type 2 (PCV-2) is an important pathogen in swine, a PCR method was established by using a set of primers derived from the published nucleotide sequence of the PCV-2 genes deposited in GenBank. In order to confirm its specificity, the PCR was performed by using the template DNA from the reference virus, and the expected products of 536 bp were only detected in PCV-2, whereas no specific band was detected in other virus such as PRV, CSFV, PPV and PK-15 cells. The lowest DNA consistence which could be detected by the PCR method was 29 ng/L, and the sequence of aim band was belong to PCV-2. To further investigate the prevalence of PCV-2, 91 clinic samples were obtained from Shanghai and circumjacent regions in 2008, and submitted to PCR detection. The data indicated that PCV-2 in 52 samples were positive, while positive proportion was 57.14%. The clinical experiments indicate the PCR method could be used in diagnosis and epidemiology investigating.