应用商业实时RT-PCR法监测猪瘟:通过检测连续组织和血液样本进行评估

M. Tignon, G. Kulcsar, K. Belák, A. Haegeman, T. Barna, K. Fábián, R. Lévai, A. Farsang, Y. Stede, R. Vrancken, F. Koenen
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引用次数: 4

摘要

在28头猪感染了一种中等毒力的野猪源经典猪瘟病毒(CSFV)毒株('11722-WIL')的实验环境中,对TaqVet CSFV实时RT-PCR检测(TaqVet CSF)作为猪瘟(CSF)群体监测诊断工具的适用性进行了评估。根据实时荧光定量pcr (rRT-PCR)测定的病毒在组织中的分布情况,发现扁桃体最适合检测猪瘟病毒。在扁桃体中,在潜伏期,早在感染后2天(dpi),以及在临床阶段直到实验结束(24 dpi)期间检测病毒基因组。在血液中,与相应的扁桃体相比,病毒RNA检测延迟了2天。在肌肉中不规律地检测到病毒,表明肉品监测猪瘟病毒的可靠性较差。对于监测方案,还对在动物组范围内收集的血液样本评估了rRT-PCR试剂盒的有效性,并将其与基于扁桃体分析的个体诊断进行了比较。本研究表明,在感染的早期阶段,使用大量血液样本或较少数量的扁桃体可以达到相当的检测效率。增加取样量是必要的,以补偿血液中观察到的较低的病毒载量。根据这些结果,可以建议使用TaqVet CSF试剂盒进行血液检测,作为一种可接受的替代扁桃体监测的方法,前提是进行扩展采样。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Application of a Commercial Real-Time RT-PCR Assay for Surveillance of Classical Swine Fever: Evaluation by Testing Sequential Tissue and Blood Samples
The applicability of the commercial TaqVet CSFV real-time RT-PCR assay (TaqVet CSF) as a diagnostic tool for herd surveillance of classical swine fever (CSF) was evaluated in an experimental setup on a group of 28 pigs infected with a moderately virulent classical swine fever virus (CSFV) strain of wild boar origin ('11722-WIL'). Based on the viral distribution determined in tissue by the real-time RT-PCR assay (rRT-PCR), the tonsils were found the most suitable tis- sue for CSFV detection. In the tonsils, the viral genome was detected during the incubation phase, as early as 2 days post infection (dpi), and during the clinical phase until the end of the experiment (24 dpi). In blood, viral RNA detection was delayed by 2 days compared to the corresponding tonsils. Virus was irregularly detected in muscles, indicating the poor reliability of meat for CSFV monitoring. For surveillance programmes, the effectiveness of the rRT-PCR kit was also evaluated on blood samples collected at the scale of the animal group and compared to the individual diagnosis based on analysis of tonsils. The present study indicated that, at the very early stages of infection, comparable detection efficiency was achieved by using large numbers of blood samples or smaller number of tonsils. The increased size of sampling is necessary to compensate the lower viral load observed in blood. Based on these results, testing of blood can be proposed as an acceptable alternative to tonsils for herd surveillance with the TaqVet CSF kit, providing that extended sampling is undertaken.
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