PRV和PCV2二重PCR检测方法的建立与初步应用

为建立同时检测猪伪狂犬病病毒(pseudorabies virus,PRV)和猪圆环病毒2型(porcine circovirus 2,PCV2)2种病毒的多重PCR,设计2对PRV和PCV2特异性引物,建立同时检测这2种病毒的二重PCR方法,并对采自呼吸障碍病猪的病料进行检测。结果表明,PRV和PCV2扩增产物分别为359bp和482bp,最小DNA质量分别是2.7pg和4.3pg。采自河南省的80份样品的总阳性率为85%(68/80),其中PRV和PCV2阳性率分别为28.8%(23/80)和77.5%(62/80),混合感染率达25%(17/68)。该多重PCR检测方法敏感、特异、快速,可用于临床样品PRV和PCV2检测。

Development and Preliminary Application of a Dulplex Polymerase Chain Reaction for Detecting Pseudorabies Virus and Porcine Circovirus 2

To simultaneously detect pseudorabies virus (PRV) and porcine circovirus type 2 (PCV2), two pairs of primers specific to PRV and PCV2 are designed and a dulplex PCR was developed. The samples from pigs with respiratory failure were detected by the dulplex PCR. The results showed that products for PRV and PCV2 are 359 bp and 482 bp, respectively. The DNA detection limit of the multiplex PCR was 2.7 pg and 4.3 pg for PRV and PCV2, respectively. Among 100 pig specimens from Henan province, the total positive rate was 85% (68/80) and within them, the positive rate for PRV and PCV2 was respectively 28.8% (23/80) and 77.5% (62/80). And the co-infection rate was 25% (17/68). This dulplex PCR is sensitive, specific and rapid method for the detection and identification of PRV and PCV2 in clinical samples.