关键词: 猪繁殖与呼吸综合征病毒, 反转录-聚合酶链反应, 检测

Abstract: According to the GenBank published sequence of porcine reproductive and respiratory sydrome virus (PRRSV), P1/P2 primes were designed to amplify the American-type gene (about 550 bp), and P3/P4 primes were designed to amplify the European-type gene (about 430 bp). In the research, we established the identifying and distinguishing PRRSV RT-PCR method with the P1/P2 and P3/P4 primes. There was no amplification observed when using RNA or DNA extracted from pseudorabies virus (PRV), Porcine parvovirus (PPV) and Classical Swine fever virus (CSF), which showed the assay had good specificity. And the assay was also highly sensitive and could detected as little as 10^-5 TCID₅₀ and 10^-4 TCID₅₀ of virus template. The lung, spleen and lymphoid tissues of 20 doubted-to-be-infected-with-PRRSV pigs from the pig farms in Chongqing and Sichuan were detected by RT-PCR. A gene fragment about 550 bp was amplified from 15 of 20 samples, while a gene fragment about 430 bp was not amplified, the results confirmed the amplified mixed tissues belonged to Americana type. The RT-PCR was proved to be a specific, sensitive, rapid and simple method and could be used to research on epidemiology and early clinical diagnosis of PRRS in molecular level.

Key words: PRRSV, RT-PCR, detection