Bird Patrick, Fisher Kiel, Boyle Megan, Huffman Travis, Benzinger M Joseph, Bedinghaus Paige, Flannery Jonathon, Crowley Erin, Agin James, Goins David, Benesh DeAnn, David John
Q Laboratories, Inc., 1400 Harrison Ave, Cincinnati, OH 45214, USA.
J AOAC Int. 2014 Sep-Oct;97(5):1329-42. doi: 10.5740/jaoacint.14-101.
The 3M(™) Molecular Detection Assay (MDA) Salmonella utilizes isothermal amplification of nucleic acid sequences with high specificity, efficiency, rapidity and bioluminescence to detect amplification of Salmonella spp. in food, food-related, and environmental samples after enrichment. A method modification and matrix extension study of the previously approved AOAC Official Method(SM) 2013.09 was conducted, and approval of the modification was received on March 20, 2014. Using an unpaired study design in a multilaboratory collaborative study, the 3M MDA Salmonella method was compared to the U.S. Department of Agriculture/Food Safety and Inspection Service (USDA/FSIS) Microbiology Laboratory Guidebook (MLG) 4.05 (2011), Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg, and Catfish Products for raw ground beef and the U.S. Food and Drug Administration (FDA)/Bacteriological Analytical Manual (BAM) Chapter 5, Salmonella reference method for wet dog food following the current AOAC guidelines. A total of 20 laboratories participated. For the 3M MDA Salmonella method, raw ground beef was analyzed using 25 g test portions, and wet dog food was analyzed using 375 g test portions. For the reference methods, 25 g test portions of each matrix were analyzed. Each matrix was artificially contaminated with Salmonella at three inoculation levels: an uninoculated control level (0 CFU/test portion), a low inoculum level (0.2-2 CFU/test portion), and a high inoculum level (2-5 CFU/test portion). In this study, 1512 unpaired replicate samples were analyzed. Statistical analysis was conducted according to the probability of detection (POD). For the low-level raw ground beef test portions, the following dLPOD (difference between the LPODs of the reference and candidate method) values with 95% confidence intervals were obtained: -0.01 (-0.14, +0.12). For the low-level wet dog food test portions, the following dLPOD with 95% confidence intervals were obtained: -0.04 (-0.16, +0.09). No significant differences were observed in the number of positive samples detected by the 3M MDA Salmonella method versus either the USDA/FSIS-MLG or FDA/BAM methods.
3M（™）分子检测分析法（MDA）沙门氏菌检测法利用核酸序列的等温扩增技术，具有高特异性、高效性、快速性和生物发光特性，用于检测富集后食品、食品相关及环境样品中的沙门氏菌属扩增情况。对先前批准的AOAC官方方法（SM）2013.09进行了方法改进和基质扩展研究，并于2014年3月20日获得了该改进方法的批准。在一项多实验室协作研究中采用非配对研究设计，将3M MDA沙门氏菌检测法与美国农业部/食品安全与检验局（USDA/FSIS）微生物学实验室指南（MLG）4.05（2011年）（用于生牛肉末中沙门氏菌的分离与鉴定）以及美国食品药品监督管理局（FDA）/细菌学分析手册（BAM）第5章（遵循当前AOAC指南的湿狗粮沙门氏菌参考方法）进行比较。共有20个实验室参与。对于3M MDA沙门氏菌检测法，生牛肉末采用25克测试份进行分析，湿狗粮采用375克测试份进行分析。对于参考方法，每种基质均采用25克测试份进行分析。每种基质分别以三个接种水平人工接种沙门氏菌：未接种对照水平（0 CFU/测试份）、低接种量水平（0.2 - 2 CFU/测试份）和高接种量水平（2 - 5 CFU/测试份）。在本研究中，共分析了1512个非配对重复样品。根据检测概率（POD）进行统计分析。对于低水平生牛肉末测试份，获得了以下具有95%置信区间的dLPOD（参考方法和候选方法的LPOD差值）值：-0.01（-0.14，+0.12）。对于低水平湿狗粮测试份，获得了以下具有95%置信区间的dLPOD值：-0.04（-0.16，+0.09）。3M MDA沙门氏菌检测法检测到的阳性样品数量与USDA/FSIS - MLG或FDA/BAM方法相比，未观察到显著差异。
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