猪蓝耳病病毒变异株TaqMan探针实时荧光定量RT-PCR鉴别诊断方法的建立

Establishment of TaqMan Probe Fluorescent RT-PCR Assay for Differential Detection of PRRSV Variant Strains

根据GenBank收录的PRRSV基因序列,设计一对引物F、R和一条TaqMan荧光探针P,建立了猪蓝耳病病毒荧光RT-PCR检测方法。该方法敏感性比常规的RT-PCR高100倍,对PRRSV细胞培养物的检测下限可以达到2个TCID50,能特异检测出蓝耳病病毒,并且可以鉴别诊断PRRSV变异株和经典株,同时根据灵敏度试验建立的标准曲线,能够对PRRSV RNA进行相对定量。

A method to detect porcine reproductive and respiratory syndrome virus （PRRSV） was developed using fluorescent RT- PCR method. Based on the published PRRSV genomic sequence in GenBank, a pair of primers F, R and a TaqMan fluorescent probe P were designed. This method was more sensitive than conventional RT-PCR by 100-fold. It can detect cell culture material of PRRSV as little as 2 TCID50. With high specificity, it could well differentiate variant and classical PRRSV strains. Besides, it could relatively quantificate PRRSV RNA according to the established standard curve of sensitivity experiment.