关键词: 猪场, 疾病, 猪繁殖与呼吸综合征, PRRSV, 双重TaqMan qPCR, 混合感染, 检测

Abstract:

In order to achieve simultaneous and rapid detection and differentiation of porcine reproductive and respiratory syndrome virus types 1 and 2, a pair of specific primers and a TaqMan MGB probe were designed with the ORF6 gene sequence of the common reference strains of PRRSV 1 and PRRSV 2 as the target sequences. By optimizing the reaction conditions and carrying out specificity, reproducibility and sensitivity tests, a dual qPCR method for the rapid identification of PRRSV 1 and PRRSV 2 was established. The results showed that the minimum detection limit of the PRRSV 1 positive plasmid standard was 10 copies/滋L, and the minimum detection limit of the PRRSV 2 positive plasmid standard was 102 copies/滋L, indicating high sensitivity. The coefficient of variation (CV) of the intra-assay and inter-assay repeatability tests of this method was below 1.4%, and the R2 of PRRSV 1 and PRRSV 2 were 0.936 and 0.985, respectively, indicating good repeatability. This method was able to specifically amplify PRRSV 1 and PRRSV 2 clinical positive samples, and had no cross-reactivity with 8 common porcine infectious pathogens such as porcine pseudorabies virus (PRV) and negative controls, with significant specificity. In summary, the PRRSV 1 and PRRSV 2 dual qPCR method established in this study could identify and detect the two pathogens at the same time, which provided technical support for the rapid detection of PRRSV and its molecular epidemiological investigation in pig farms.

Key words: pig farm')">

pig farm, disease, PRRS, PRRSV, dual TaqMan qPCR, co-infection, test