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Comparison of Sample Types and Analytical Methods for the Detection of Highly Campylobacter-Colonized Broiler Flocks at Different Stages in the Poultry Meat Production Chain

机译:在禽肉生产链中不同阶段检测高度弯曲杆菌定殖的肉鸡群的样品类型和分析方法的比较

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Exclusion of broiler batches, highly colonized with Campylobacter (>7.5 log(10) colony-forming units/g), from the fresh poultry meat market might decrease the risk of human campylobacteriosis. The objective of this study was to compare different sample types (both at the farm and the slaughterhouse) and methods (direct culture, quantitative real-time polymerase chain reaction [qPCR], propidium monoazide [PMA]-qPCR) applied for the quantification of the Campylobacter colonization level. In addition, the applicability of the lateral flow-based immunoassay, Singlepath((R)) Direct Campy Poultry test (Singlepath((R)) test), was evaluated as a rapid method for the qualitative detection of Campylobacter in highly colonized broiler batches. Campylobacter counts differed significantly between sample types collected at farm level (cecal droppings, feces, boot swabs) and at slaughterhouse level (cecal content, fecal material from crates). Furthermore, comparison of Campylobacter counts obtained by different methods (direct culture, qPCR, PMA-qPCR) in cecal droppings revealed significant differences, although this was not observed for cecal-content samples. Evaluation of the Singlepath((R)) test on cecal droppings and cecal-content samples revealed an acceptable level of sensitivity and specificity. In conclusion, cecal droppings and cecal content are proposed as the most representative sample types for quantification of Campylobacter colonization level of broilers at farm and slaughterhouse, respectively. Direct culture and qPCR are equally sensitive for quantification of Campylobacter in fresh cecal-content samples. PMA treatment before qPCR inhibits the signal from dead Campylobacter cells. Consequently, when samples are extensively stored and/or transported, qPCR is preferred to direct culture and PMA-qPCR. Furthermore, the Singlepath((R)) test offers a convenient alternative method for rapid detection of Campylobacter in highly colonized broiler batches.
机译:从新鲜的家禽肉类市场中排除高度定居于弯曲杆菌属(> 7.5 log(10)菌落形成单位/ g)的肉鸡批次可能会降低人类弯曲菌病的风险。这项研究的目的是比较不同样品类型(包括农场和屠宰场)和方法(直接培养,定量实时聚合酶链反应[qPCR],单叠氮化丙锭[PMA] -qPCR)用于定量弯曲杆菌定植水平。此外,评估了侧向流式免疫测定的适用性,即定性检测高度定植的肉鸡批次中弯曲杆菌的一种快速方法,该方法直接定性检测了Campy Poultry家禽测试(Singlepath(R))。 。在农场水平(粪便,粪便,靴子拭子)和屠宰场水平(粪便含量,来自板条箱的粪便)收集的样本类型之间,弯曲杆菌计数存在显着差异。此外,盲肠粪便中通过不同方法(直接培养,qPCR,PMA-qPCR)获得的弯曲杆菌计数比较显示出显着差异,尽管盲肠含量样品未观察到。对盲肠粪便和盲肠内容物样品的Singlepath(R)测试的评估显示出可接受的敏感性和特异性水平。总之,盲肠粪便和盲肠含量被认为是量化农场和屠宰场肉鸡弯曲杆菌定殖水平最有代表性的样品类型。直接培养和qPCR对新鲜盲肠含量样品中的弯曲杆菌定量同样敏感。 qPCR之前的PMA处理可抑制死去的弯曲杆菌细胞的信号。因此,当样品被大量存储和/或运输时,qPCR优于直接培养和PMA-qPCR。此外,Singlepath(R)测试提供了一种方便的替代方法,用于快速检测高度定居的肉鸡批次中的弯曲杆菌。

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