猪主要繁殖障碍疾病病原的多重实时定量PCR快速检测

Abstract

Abstract:

The aim of this study is to establish a multiplex real-time PCR assay to simultaneously detect and discriminate porcine reproductive and respiratory syndrome virus (PRRSV)，porcine circovirus type 2 (PCV2)，porcine pseudorabies virus（PRV）and porcine parvovirus (PPV) in a single test tube. According to the nucleotide sequences of PRRSV，PCV2, PRV and PPV from GenBank, four pairs of specific probes were designed. By optimizing the probe’s concentration, primers concentration and sample DNA extraction, a multiplex real-time quantitative PCR assay for detection of these viruses was established. The specificity and sensitivity of the multiplex real-time quantitative PCR assay were analyzed for single virus and multiple viruses. The results showed that the specificity was high, and no amplification was achieved to the other pathogeneses. Both multiplex and singleplex assays were consistently able to detect ＜10¹ copies of plasmid templates or 1 TCID₅₀ virus per reaction. 40 clinical samples were comparatively detected using the multiplex and singleplex assay. The data showed that the results of the singleplex and multiplex assay were in accordance with the results of conventional PCR. A multiplex real-time PCR assay was developed to simultaneously detect and discriminate PRRSV, PCV2, PRV and PPV in a single test tube, the method is rapid, sensitive, specific and accurate.

CLC Number:

S852.285.3

ZHAO Xu-yong, ZHAO Li, NING Yu-chang, MA Hui. Simultaneous Detection of Major Pathogens of Reproductive Failure Diseases in Swine by a Multiplex Real-time PCR[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, doi: .

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Simultaneous Detection of Major Pathogens of Reproductive Failure Diseases in Swine by a Multiplex Real-time PCR

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