Dipartimento di Valorizzazione e Protezione delle Risorse Agroforestali, Facoltà di Agraria, Università di Torino, Turin, Italy.
Int J Food Microbiol. 2010 Jul 31;141 Suppl 1:S75-9. doi: 10.1016/j.ijfoodmicro.2010.02.002. Epub 2010 Feb 12.
Campylobacter-contaminated food products are currently the cause of the highest number of gastroenteritis cases in developed countries. Apart for biosafety measures at the primary production level, no other official control measures are currently in place for its control. This is partly due to the lack of quantitative data regarding the prevalence and contamination level of different food products by Campylobacter spp. that does not allow for quantitative risk assessment. PCR-based methods, applied without prior enrichment, in food samples circumvent limitations associated with the quantification of foodborne pathogens by traditional, culture-dependent methods. In this study, we report the development of a protocol, based on the amplification of the rpoB gene of Campylobacter jejuni, by quantitative PCR (qPCR), directly in food samples. The quantification limit of the protocol was determined to be in the order of 10 colony forming units (cfu)/g or ml of food sample. The optimized protocol was applied for the survey of C. jejuni in naturally contaminated poultry samples. In parallel, traditional sampling was also performed. A high percentage of samples (87%) resulted to be positive by qPCR, while no C. jejuni was detected by traditional analysis. Furthermore, important differences were observed in the detection by qPCR between samples before and after enrichment.
受弯曲菌污染的食品目前是发达国家引发食源性胃肠炎病例的首要病因。除了在初级生产阶段采取生物安全措施外,目前尚无其他官方控制措施来控制弯曲菌。这在一定程度上是由于缺乏关于不同食品中弯曲菌属的流行程度和污染水平的定量数据,从而无法进行定量风险评估。基于聚合酶链反应(PCR)的方法,无需事先进行富集,即可应用于食品样本,避免了传统、基于培养的方法定量检测食源性致病菌所存在的局限性。在本研究中,我们报告了一种基于禽弯曲菌 rpoB 基因的定量 PCR(qPCR)扩增的方案,该方案可直接在食品样本中进行。该方案的定量限确定为每克或毫升食品样本中约 10 个菌落形成单位(cfu)。该优化方案应用于自然污染禽肉样本中弯曲菌属的检测。同时,还进行了传统采样。qPCR 检测结果显示,有很高比例的样本(87%)呈阳性,而传统分析则未检测到弯曲菌。此外,qPCR 检测结果与富集前后的样本之间存在重要差异。