Tebbs Robert S, Balachandran Priya, Wong Lily Y, Zoder Patrick, Furtado Manohar R, Petrauskene Olga V, Cao Yanxiang
Life Technologies Corp., 850 Lincoln Centre Dr, Foster City, CA 94404, USA.
J AOAC Int. 2011 Sep-Oct;94(5):1481-9. doi: 10.5740/jaoacint.11-052.
Increasingly, more food companies are relying on molecular methods, such as PCR, for pathogen detection due to their improved simplicity, sensitivity, and rapid time to results. This report describes the validation of a new Real-Time PCR method to detect Listeria monocytogenes in nine different food matrixes. The complete system consists of the MicroSEQ L. monocytogenes Detection Kit, sample preparation, the Applied Biosystems 7500 Fast Real-Time PCR instrument, and RapidFinder Express software. Two sample preparation methods were validated: the PrepSEQ Nucleic Acid extraction kit and the PrepSEQ Rapid Spin sample preparation kit. The test method was compared to the ISO 11290-1 reference method using an unpaired-study design to detect L. monocytogenes in roast beef, cured bacon, lox (smoked salmon), lettuce, whole cow's milk, dry infant formula, ice cream, salad dressing, and mayonnaise. The MicroSEQ L. monocytogenes Detection Kit and the ISO 11290-1 reference method showed equivalent detection based on Chi-square analysis for all food matrixes when the samples were prepared using either of the two sample preparation methods. An independent validation confirmed these findings on smoked salmon and whole cow's milk. The MicroSEQ kit detected all 50 L. monocytogenes strains tested, and none of the 30 nontargeted bacteria strains.
越来越多的食品公司开始依赖分子方法，如聚合酶链反应（PCR）来进行病原体检测，因为这些方法操作更简便、灵敏度更高且出结果更快。本报告描述了一种用于检测九种不同食品基质中单核细胞增生李斯特菌的新型实时PCR方法的验证情况。完整系统包括MicroSEQ单核细胞增生李斯特菌检测试剂盒、样品制备、Applied Biosystems 7500快速实时PCR仪器以及RapidFinder Express软件。验证了两种样品制备方法：PrepSEQ核酸提取试剂盒和PrepSEQ快速离心样品制备试剂盒。采用非配对研究设计，将该测试方法与ISO 11290-1参考方法进行比较，以检测烤牛肉、腌制培根、熏鲑鱼（lox）、生菜、全脂牛奶、婴儿配方奶粉、冰淇淋、沙拉酱和蛋黄酱中的单核细胞增生李斯特菌。当使用两种样品制备方法中的任何一种制备样品时，基于卡方分析，MicroSEQ单核细胞增生李斯特菌检测试剂盒和ISO 11290-1参考方法在所有食品基质中均显示出等效的检测效果。一项独立验证在熏鲑鱼和全脂牛奶上证实了这些结果。MicroSEQ试剂盒检测了所有50株测试的单核细胞增生李斯特菌菌株，而30株非目标细菌菌株均未被检测到。
