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A Duplex Real-Time PCR Assay for the Simultaneous Detection of Porcine Circovirus 2 and Circovirus 3

机译:同时检测猪圆环病毒2和圆环病毒3的双重实时PCR检测

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摘要

Porcine circoviruses (PCV) include PCV1, PCV2, and the new-emerging PCV3. PCV2 is pathogenic to pigs, but the pathogenicity of PCV3 in pigs is debatable. Recently, there have been frequent reports of PCV2 and PCV3 co-infections in clinical samples. Thus, it would be practical to develop a duplex PCR method to detect PCV2 and PCV3 simultaneously. In this study, specific primers and probes were designed to target PCV2 cap and PCV3 rep genes. A duplex real-time PCR method was then developed to detect the two viruses. The assay was found to be highly specific, sensitive, and reproducible for PCV2/3 without cross-reactions with other swine pathogens. The sensitivity of this assay was 2.9 copies for the PCV2 plasmid and 22.5 copies for the PCV3 plasmid. The established assay was then used to detect PCV2/3 infection in 340 clinical samples collected in the first half of 2017. The results showed that the co-infection rate of PCV2/3 in the samples was 27.6%. Our study provides an important tool that can be used to perform urgently needed surveys for the two porcine circoviruses to evaluate their impact on the swine industry.Electronic supplementary materialThe online version of this article (10.1007/s12250-018-0025-2) contains supplementary material, which is available to authorized users.
机译:猪圆环病毒(PCV)包括PCV1,PCV2和新出现的PCV3。 PCV2对猪具有致病性,但猪中PCV3的致病性尚有争议。最近,在临床样品中经常有PCV2和PCV3合并感染的报道。因此,开发一种同时检测PCV2和PCV3的双链PCR方法将是可行的。在这项研究中,设计了针对PCV2 cap和PCV3 rep基因的特异性引物和探针。然后开发了一种双重实时PCR方法来检测这两种病毒。发现该测定法对PCV2 / 3具有高度特异性,敏感性和可重复性,并且不会与其他猪病原体发生交叉反应。该测定的敏感性对于PCV2质粒为2.9拷贝,对于PCV3质粒为22.5拷贝。然后,使用已建立的分析方法检测2017年上半年收集的340份临床样品中的PCV2 / 3感染。结果表明,样品中PCV2 / 3的共感染率为27.6%。我们的研究提供了一种重要工具,可用于对两种猪圆环病毒进行急需的调查,以评估其对猪业的影响。电子补充材料本文的在线版本(10.1007 / s12250-018-0025-2)包含补充信息资料,可供授权用户使用。

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