PCR技术在猪圆环病毒2型检测中的应用

The Application of PCR Method to Detect Porcine circovirus Ⅱ

根据猪圆环病毒2型(PCV-2)ORF2基因序列,设计并合成了一对引物,以本室分离鉴定的PCV-2为模板,筛选最佳反应条件,建立了检测PCV-2的PCR方法。应用该方法对PCV-2进行基因扩增,获得长度为559 bp的特异性目的DNA片段,而对PRRSV、CSFV、PPV等病毒进行同条件检测,结果均为阴性,无交叉反应;敏感性试验表明,该体系可检测到10.8 pg的PCV-2基因组DNA。对2004年-2005年期间送检的81份临床样品进行检测,阳性率为22.2%。表明所建立的PCR技术可用于PCV感染的诊断和流行病学调查。

The optimization PCR method to detect the Porcine Circovirus type Ⅱ （PCV-2） were established after a pair of primer were designed and synthesized based on the sequences of the PCV-2 genome. This PCR technique was applied to specifically amplify the 559 bp DNA fragment of the PCV-2. The negative results were achieved from porcine reproductive and respiratory syndrome virus（PRRSV）, Classical swine fever virus（CSFV）, Porcine parvovirus（PPV） and PK-15 cell. The blast of the gene sequence of PCR product achieved to 100% homology with that of other PCV-2 strains. The sensitivity of PCR reached to 10.8 pg of PCV-2 DNA. During the 2004-2005, the positive ratio of 81 clinical detected specimen were reached to 22.2%. The results showed that this PCR technique was specific and applied to PCV-2 diagnoses and epidemiology investigation.