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Detection of Food Hazards in Foods: Comparison of Real Time Polymerase Chain Reaction and Cultural Methods

机译:检测食品中的食品危害:实时聚合酶链反应和培养方法的比较

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摘要

Foodstuffs should not contain microorganisms or their toxins or metabolites in quantities suggesting an unacceptable risk for human health. The detection of food hazards in foods is performed by several tests that produce results dependent on the analytical method used: an analytical reference method, defined as standard, is associated with each microbiological criterion laid down in Regulation 2073/2005/EC, but, analytical methods other than the reference ones, in particular more rapid methods, could be used. Combined screening methods performed by real time-polymerase chain reaction (RT-PCR) are currently validated as alternative methods according to the ISO 16140:2003 and certified by the Association Française de Normalisation. However, the positive results obtained with these alternative methods, the investigated molecular relations that resulted positive have to be confirmed with cultural methods using the same enrichment media in which the molecular screening was performed. Since it is necessary to assess if these testing schemes provide equivalent guarantees of food safety, the aim of this retrospective study is to analyse the data collected, from 2012 to 2014 by Emilia Romagna Region in the field of Piano Regionale Alimenti (Food Regional Plan) during official controls monitoring food samples of animal and other than animal origin. Records performed by combined methods of molecular screening of Salmonella spp., Listeria monocytogenes and thermophilic Campylobacter and cultural confirmation results were gathered together and the results were compared in order to assess the sensitivity of the methods. A total of 10,604 food samples were considered in this study: the comparison of the data revealed that the RT-PCR method detected Salmonella, L. monocytogenes, and thermophilic Campylobacter in 2.18, 3.85 and 3.73% of the samples, respectively, whereas by using cultural method these pathogens were isolated in 0.43, 1.57 and 1.57% of samples, respectively. In spite of the use of the same enrichment broth, the RT-PCR method disclosed a percentage of positive samples that was negative to cultural examination ranging between 20 and 43%, with a PCR/culture ratio between 2.37 to 5.00. In conclusion, the results of this study pose a doubt about the sensitivity of the official cultural methods regarding the isolation of the three investigated foodborne pathogens. Moreover this study may be a useful tool for veterinary authorities to assess appropriate sampling plans to control the risk relating to the consumption of contaminated foods.
机译:食品中不得含有表明人体健康风险不可接受的微生物或其毒素或代谢产物。食品中食品危害的检测通过多种测试来进行,这些测试的结果取决于所使用的分析方法:定义为标准的分析参考方法与法规2073/2005 / EC中规定的每个微生物标准相关联,但是,可以使用参考文献以外的方法,尤其是更快速的方法。目前,通过实时聚合酶链反应(RT-PCR)进行的联合筛选方法已根据ISO 16140:2003进行了验证,并已通过法国标准化协会认证。但是,通过这些替代方法获得的阳性结果,必须使用与进行分子筛查相同的富集培养基的培养方法,确认所研究的导致阳性的分子关系。由于有必要评估这些测试方案是否能够提供等同的食品安全保证,因此,本回顾性研究的目的是分析艾米利亚·罗马涅地区从2012年至2014年在Piano Regionale Alimenti(食品区域计划)领域收集的数据在官方控制期间,监测动物和非动物来源的食物样本。通过沙门氏菌,单核细胞增生性李斯特菌和嗜热弯曲杆菌的分子筛查联合方法进行的记录和培养确认结果汇总在一起,并对结果进行比较,以评估方法的敏感性。本研究共考虑了10,604个食品样品:数据比较表明,RT-PCR方法分别在样品中分别检出了沙门氏菌,单核细胞增生李斯特菌和嗜热弯曲杆菌,分别占2.18%,3.85%和3.73%。培养方法这些病原体分别在0.43、1.57和1.57%的样品中分离。尽管使用了相同的富集肉汤,但RT-PCR方法显示对培养物检查呈阴性的阳性样品百分比在20%至43%之间,PCR /培养物比率在2.37至5.00之间。总之,这项研究的结果使人们怀疑官方文化方法对分离三种被调查的食源性病原体的敏感性。此外,这项研究可能是兽医当局评估适当抽样计划以控制与食用污染食品有关的风险的有用工具。

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