Jaroenporn Chanokchon, Supawasit Wannakarn, Bundidamorn Damkerng, Udompijitkul Pathima, Assawamakin Anunchai, Trevanich Sudsai
Department of Food Science and Technology, Faculty of Agro-Industry, Kasetsart University, Bangkok 10900, Thailand.
Center for Advanced Studies for Agriculture and Food, Institute for Advanced Studies, Kasetsart University, Bangkok 10900, Thailand.
Foods. 2022 May 25;11(11):1557. doi: 10.3390/foods11111557.
The aim of the study was to perform in-house validation of the developed multiplex PCR (mPCR)-based alternative method to detect Shiga toxin-producing (STEC), () and spp. in raw meats following the ISO 16140-2: 2016. A comparative study of the developed mPCR against the Bacteriological Analytical Manual (BAM) method was evaluated for inclusivity and exclusivity, sensitivity and the relative level of detection (RLOD). Inclusivity levels for each target bacterium were all 100%, while exclusivity for non-target bacteria was 100%. The sensitivity of the developed mPCR was calculated based on the analysis of 72 samples of raw meat. The sensitivity of the developed mPCR was 100%. The RLOD values of the developed mPCR for STEC, and spp. were 0.756, 1.170 and 1.000, respectively. The developed mPCR showed potential as a tool for the fast, specific and sensitive detection of the three bacteria in the raw meat industry.
本研究的目的是按照ISO 16140-2:2016标准,对所开发的基于多重聚合酶链反应(mPCR)的替代方法进行内部验证,以检测生肉中的产志贺毒素大肠杆菌(STEC)、(此处原文括号内容缺失)和(此处原文缺失具体菌名)菌属。针对所开发的mPCR与《细菌学分析手册》(BAM)方法进行了一项比较研究,评估其包容性和排他性、灵敏度以及相对检测水平(RLOD)。每种目标细菌的包容性水平均为100%,而非目标细菌的排他性为100%。基于对72份生肉样本的分析计算了所开发mPCR的灵敏度。所开发mPCR的灵敏度为100%。所开发mPCR对STEC、(此处原文缺失具体菌名)和(此处原文缺失具体菌名)菌属的RLOD值分别为0.756、1.170和1.000。所开发的mPCR显示出作为一种工具用于生肉行业中快速、特异性和灵敏检测这三种细菌的潜力。