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首页> 外文期刊>Journal of Clinical Microbiology >Single-Tube Multiplexed Molecular Detection of Endemic Porcine Viruses in Combination with Background Screening for Transboundary Diseases
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Single-Tube Multiplexed Molecular Detection of Endemic Porcine Viruses in Combination with Background Screening for Transboundary Diseases

机译：结合地方病跨境疾病背景筛查的单管复用地方性猪病毒分子检测

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Detection of several pathogens with multiplexed real-time quantitative PCR (qPCR) assays in a one-step setup allows the simultaneous detection of two endemic porcine and four different selected transboundary viruses. Reverse transcription (RT)-qPCR systems for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2), two of the most economically important pathogens of swine worldwide, were combined with a screening system for diseases notifiable to the World Organization of Animal Health, namely, classical and African swine fever, foot-and-mouth disease, and Aujeszky's disease. Background screening was implemented using the identical fluorophore for all four different RT-qPCR assays. The novel multiplex RT-qPCR system was validated with a large panel of different body fluids and tissues from pigs and other animal species. Both reference samples and clinical specimens were used for a complete evaluation. It could be demonstrated that a highly sensitive and specific parallel detection of the different viruses was possible. The assays for the notifiable diseases were even not affected by the simultaneous amplification of very high loads of PRRSV- and PCV2-specific sequences. The novel broad-spectrum multiplex assay allows in a unique form the routine investigation for endemic porcine pathogens with exclusion diagnostics of the most important transboundary diseases in samples from pigs with unspecific clinical signs, such as fever or hemorrhages. The new system could significantly improve early detection of the most important notifiable diseases of swine and could lead to a new approach in syndromic surveillance.
机译：在一步设置中使用多重实时定量PCR（qPCR）分析检测多种病原体，可以同时检测两种地方性猪和四种不同的选定跨界病毒。用于检测猪繁殖与呼吸综合征病毒（PRRSV）和2型猪圆环病毒（PCV2）的逆转录（RT）-qPCR系统与世界上最重要的猪病原体中的两种病原体相结合，可用于应报告的疾病筛查系统致世界动物卫生组织，即古典和非洲猪瘟，口蹄疫和奥吉斯基病。对于所有四个不同的RT-qPCR分析，使用相同的荧光团进行背景筛选。大量来自猪和其他动物物种的不同体液和组织对新型多重RT-qPCR系统进行了验证。参考样品和临床标本均用于完整评估。可以证明，对不同病毒进行高度灵敏和特异性的平行检测是可能的。可以同时扩增非常高负荷的PRRSV和PCV2特异性序列的方法，甚至不影响应报告疾病的检测。新颖的广谱多重测定法以独特的形式允许对地方性猪病原体进行常规调查，并排除具有特殊临床症状（如发烧或出血）的猪样品中最重要的跨界疾病。新系统可以显着改善对最重要的应通报猪病的早期发现，并可能导致新的综合征监测方法。

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