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首页> 外文期刊>Food Analytical Methods >Validation of Real-Time PCR and Enzyme-Linked Fluorescent Assay-Based Methods for Detection of Salmonella spp. in Chicken Feces Samples
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Validation of Real-Time PCR and Enzyme-Linked Fluorescent Assay-Based Methods for Detection of Salmonella spp. in Chicken Feces Samples

机译：实时荧光定量PCR和基于酶联荧光法的沙门氏菌检测方法的验证。在鸡粪样品中

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摘要

A comparative study of enzyme-linked fluorescent assay (ELFA)-based methods and real-time polymerase chain reaction (PCR)-based methods using three and two different sample preparation protocols, respectively, and the standard culture-based method EN ISO 6579:2002/Amd1:2007, for the detection of Salmonella spp. in chicken feces, was performed on 20 artificially and 68 naturally contaminated chicken feces. Selectivity, relative specificity, relative accuracy, relative sensitivity, and relative detection level were determined. According to criteria established in the methods comparison study included in EN ISO 16140:2003 for validation of alternative microbiological methods, the ELFA-based methods (V1 and V2) as well as a real-time PCR method (PCR2) were comparable to the reference method for the detection of Salmonella in chicken feces. They provided results in 48 h and presented a high sensitivity (97% for all of them). The three methods showed a relative specificity of 94%, V1 being the method which presented the highest relative accuracy (96%). While detection level for V2 and reference method was between 3 and 13 CFU/25 g, PCR2 method was able to detect down to 3 CFU/25 g. In conclusion, both the real-time PCR and the ELFA-based assays can be used as rapid and user-friendly screening methods for detection of Salmonella spp. in chicken feces.
机译：分别使用三种和两种不同的样品制备方案以及基于标准培养物的方法EN ISO 6579对基于酶联荧光测定（ELFA）的方法和基于实时聚合酶链反应（PCR）的方法进行的比较研究： 2002 / Amd1：2007，用于检测沙门氏菌。在鸡粪中进行了人工处理，并对20种自然污染的鸡粪进行了处理。确定了选择性，相对特异性，相对准确度，相对灵敏度和相对检测水平。根据EN ISO 16140：2003中方法比较研究中确立的用于验证替代微生物方法的标准，基于ELFA的方法（V1和V2）以及实时PCR方法（PCR2）与参考文献相当鸡粪中沙门氏菌的检测方法。他们在48小时内提供了结果，并表现出很高的敏感性（所有患者的97％）。三种方法的相对特异性为94％，V1是相对准确度最高的方法（96％）。 V2和参考方法的检测水平在3到13 CFU / 25 g之间，而PCR2方法只能检测到3 CFU / 25 g。总之，实时荧光定量PCR和基于ELFA的测定均可用作检测沙门氏菌的快速且用户友好的筛选方法。在鸡粪里。

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