Hubei Key Laboratory of Animal Embryo and Molecular Breeding, Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, PR China.
Gene. 2013 Dec 1;531(2):199-204. doi: 10.1016/j.gene.2013.09.017. Epub 2013 Sep 11.
Outbreaks of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) have led to large economic losses and, subsequently, have drawn great attention to its diagnosis and prevention. To facilitate rapid discrimination of HP-PRRSV from classical PRRSV (C-PRRSV), we developed a one-step RT-PCR assay. Primer specificities were evaluated with RNA extracted from 8 viral strains and our results revealed that the primers had a high specificity for PRRSV. The assay sensitivity was 25 copies/μL for both HP-PRRSV and C-PRRSV. A total of 929 serum samples were identified, of which 20.45% were HP-PRRSV-positive and 1.51% were C-PRRSV-positive, which was completely consistent with that of immunochromatochemistry and sequencing method. The proposed assay can detect the virus 2 days prior the onset of symptoms and it can be performed in 2h, thereby providing a rapid method to discriminate HP-PRRSV from C-PRRSV for the identification and prevention of PRRSV infections.
高致病性猪繁殖与呼吸综合征病毒（HP-PRRSV）的爆发导致了巨大的经济损失，因此引起了人们对其诊断和预防的极大关注。为了方便快速区分 HP-PRRSV 和经典 PRRSV（C-PRRSV），我们开发了一种一步 RT-PCR 检测方法。用从 8 株病毒中提取的 RNA 评估了引物的特异性，结果表明这些引物对 PRRSV 具有高度特异性。该检测方法对 HP-PRRSV 和 C-PRRSV 的灵敏度均为 25 拷贝/μL。共鉴定了 929 份血清样本，其中 20.45%为 HP-PRRSV 阳性，1.51%为 C-PRRSV 阳性，与免疫层析化学法和测序法完全一致。该检测方法可在症状出现前 2 天检测到病毒，且可在 2 小时内完成，因此为鉴别 PRRSV 感染中的 HP-PRRSV 和 C-PRRSV 提供了一种快速方法。
