关键词: 猪繁殖与呼吸综合征病毒, 实时荧光定量RT-PCR, 一步法

Abstract: To identify porcine reproductive and syndrome virus (PRRSV) rapidly using realtime RT-PCR technique, primers and TaqMan probe were designed based on the PRRSV sequence. The fluorescent quantitative RT-PCR assay for detection of PRRSV was established. Its specificity, sensitivity and stability were examined. The assay was also proven to be specific based on the results that no amplification was found by detecting RNA/DNA from PCV, JEV, SFV, PRV and PPV. 10 copies of template RNA or one TCID50 of PRRSV could be detected. The coefficient variation (CV%) of intra/interassay for same DNA sample was less than 1%. 72 samples were examined by the fluorescent quantitative RT-PCR and the conventional RTPCR respectively. 40 were detected to be infected with PRRSV by the realtime PCR, but only 28 were confirmed to be infected with PRRSV by conventional PCR. These results showed that the fluorescent quantitative RT-PCR assay was rapid, specific, sensitive and simple for the detection of PRRSV．

Key words: porcine reproductory syndrome virus (PRRSV), realtime fluorescent quantitative RT-PCR assay, onestep method