猪细小病毒、伪狂犬病病毒和猪圆环病毒2型多重PCR检测方法的建立及应用

Establishment and Application of a Multiplex PCR for Detecting Porcine Parvovirus,Pseudorabies Virus and Porcine circovirus type2

根据猪细小病毒(PPV)、伪狂犬病病毒(PRV)和猪圆环病毒2型(PCV-2)的基因序列,分别选取各自的保守区段设计引物,通过反应条件的优化,建立了检测PPV、PRV和PCV-2的多重PCR方法。用建立的方法对采自陕西省部分猪场的286份病料及血样进行检测,从对临床健康猪全血样品中PPV、PRV和PCV-2的检测结果看,PCV-2在正常猪群中有一定比例的存在,同时在个别猪场也存在PRV和PCV-2双重感染的现象;从临床患病猪病料中检测发现,PRV和PCV-2混合感染较严重,并有PPV+PRV+PCV-2三重感染发生;多重PCR检测结果与单项PCR检测结果的符合率达99%以上,说明建立的多重PCR检测方法可用于临床样品的检测,为疾病诊断提供参考。

A multiplex polymerase chain reaction （PCR）-based assay was developed and evaluated for its ability to simultaneously detect multiple viral infectlons of swine. Specific primers for detecting porcine parvovirus （PPV）, pseudorabies virus （PRV） and porcine circovirus type 2 （PCV-2） were designed according to the full sequences of each virus strain obtained from GenBank. A multiplex PCR was established to detect PPV, PRV and PCV-2 by optimizing the PCR programs. 286 blood and tissues samples collected in clinical healthy pig population and disease pigs of pig farms in Shaanxi were detected using developed mul- tiplex PCR for screening PPV, PRV and PCV-2. The results indicated that the PCV-2 was an important pathogen in clinical healthy pig population, the PRV and PPV also could be found in individual pig farms, so it was important issue for getting rid of the infected pigs from normal pig population. It was serous of PRV＋PCV-2 combined infection in diseased pigs, PPV＋PRV＋PCV-2 combined infection also were detected in diseased pigs. The detected results of multiplex PCR were more than 99 N consistent with special single PCR. This developed novel multiplex PCR will be a tool for detecting PPV,PRV and PCV-2 in clinical samples and disease diagnosis.