猪繁殖与呼吸综合征病毒和猪圆环病毒2型双重荧光PCR检测方法的建立

Development of a duplex real-time PCR for the detection of porcine reproductive and respiratory syndrome virus and porcine circovirus type 2

为建立快速简便检测猪繁殖与呼吸综合征病毒(PRRSV)和猪圆环病毒2型(PCV2)的方法,本研究根据GenBank中已登录的PRRSVORF7基因序列和PCV2 ORF1基因序列,设计了2对特异性引物和2条TaqMan探针,通过优化反应条件,建立了检测PRRSV和PCV2的双重荧光PCR方法。结果显示,该方法特异性好,不与其他常见猪病病原体发生交叉反应,检测PRRSV和PCV2的灵敏度分别可达1.05 TCID50/100L和6.30 TCID50/100μL,表明,本试验建立的双重TaqMan荧光定量PCR检测方法可对PRRSV和PCV2进行快速诊断,适合现场检测,为这两种疫病的诊断和防控奠定了基础。

In order to detect porcine reproductive and respiratory syndrome virus(PRRSV) and porcine circovirus type 2(PCV2),the Taq Man duplex real-time PCR was established with 2 pairs of primers and 2 specific probes of PRRSV and PCV2 designed by the ORF7 gene of PRRSV and ORF1 gene of PCV2. The results showed that it does not cross-react with the other common porcine disease pathogens for its better specificity,and its detection limit was 1.05 TCID50/100 μL of PRRSV and 6.30 TCID50/100 μL of PCV2 respectively.Furthemore,the method can be used for rapid diagnosis of PRRSV and PCV2 in the field.