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Establishment of a sensitive TaqMan‐based real‐time PCR assay for porcine circovirus type 3 and its application in retrospective quarantine of imported boars to China

机译:基于TaqMan的3型猪圆环病毒敏感实时荧光PCR检测方法的建立及其在中国进口公猪追溯检疫中的应用

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摘要

Porcine circovirus type 3 (PCV3) is a novel pathogen first identified in the United States in 2016. As there is a high possibility that no clinical signs of infection are observed in the host, an accurate and sensitive method is needed for quarantine on numerous live pigs especially for international pig trade. In this study, a TaqMan‐based real‐time PCR assay specifically for PCV3 was established without cross‐reactions with other non‐targeted pig viruses. The sensitivity of the current approach is about 1.5 × 10copies μL−1 plasmid DNA while the sensitivity of the conventional PCR is about 1.5 × 102 copies μL−1 plasmid DNA. Further, this assay was applied in the retrospective quarantine on serum samples of 601 commercial live boars imported to China from the United States, France and the United Kingdom from 2011 to 2017. The results revealed that PCV3 could be detected positive in the commercial boars imported from the United States and the above‐mentioned western European countries and phylogenetic study also revealed that viral isolates were grouped with some isolates from Korea and the United States. Our study suggested that PCV3 may be prevalent globally since 2011.
机译:猪圆环病毒3型(PCV3)是2016年在美国首次发现的新型病原体。由于极有可能在宿主中未观察到感染的临床迹象,因此需要一种准确而灵敏的方法来隔离许多活菌猪,特别是国际猪贸易。在这项研究中,建立了专门针对PCV3的基于TaqMan的实时PCR检测方法,且未与其他非靶向猪病毒发生交叉反应。当前方法的灵敏度约为1.5×10 1 拷贝μL -1 质粒DNA,而常规PCR的灵敏度约为1.5×10 2 复制μL -1 质粒DNA。此外,该检测方法用于2011年至2017年从美国,法国和英国进口到中国的601只商业活公猪的血清样品的追溯检疫中。结果表明,在进口的商业公猪中可以检测到PCV3阳性来自美国和上述西欧国家的病毒,系统发育研究还显示,病毒分离株与来自韩国和美国的一些分离株分组。我们的研究表明,自2011年以来,PCV3可能在全球范围内流行。

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