多重PCR检测猪伪狂犬病病毒、猪繁殖与呼吸综合征病毒和猪流感病毒

Detection of Pseudorabies virus,Porcine reproductive and respiratory syndrome virus and Swine influenza virus by Multiplex PCR

应用Primer3.0和Omega2.0,根据猪伪狂犬病毒(PRV)、猪繁殖与呼吸综合征病毒(PRRSV)和猪流感病毒(SIV)保守基因设计了3对多重PCR引物,建立PRRSV,SIV和PRV单项PCR检测方法,并在优化单项PCR反应条件(引物浓度、Mg2+浓度、退火温度等)基础上,初步建立了PRRSV-PRV-SIV多重PCR检测方法,并分别用多重PCR和单项PCR/RT-PCR检测10份临床病料,两者符合率为96.6%,表明该多重PCR检测方法有较高的敏感度,可以用于临床病料的检测。

Three pairs of primers were designed and analyzed with Primer 3.0 and Omega 2.0 software according to the conservative sequence M,N gene for Porcine reproductive and respiratory syndrome virus （PRRSV）, M2 gene for Swine influenza virus （SIV）,and gD gene for Pseudorabies virus（PRV）, respectively. The single PCR/RT-PCR conditions of the three viruses were optimized by determining the concentration of Mg^2＋ and the primer, and annealing temperature,et al. Based on the optimized single PCR conditions, the multiplex PCR PRRSV-PRV-SIV was developed for detection of swine respiratory disease. The 10 clinical samples were identified with the multiplex PCR and single PCR, respectively. The sensitivities of the multiplex PCR and single PCR were compared. The results indicated that the optimal conditions of the multiplex PCR layed a good foundattion of sensitive, special and rapid assay for detection of clinical samples.