Petrauskene Olga V, Cao Yanxiang, Zoder Patrick, Wong Lily Y, Balachandran Priya, Furtado Manohar R, Tebbs Robert S
Life Technologies Corp., 850 Lincoln Centre Dr, Foster City, CA 94404, USA.
J AOAC Int. 2012 Jul-Aug;95(4):1074-83. doi: 10.5740/jaoacint.11-311.
A complete system for real-time PCR detection of Listeria species was validated in five food matrixes and five environmental surfaces, namely, hot dogs, roast beef, lox (smoked salmon), pasteurized whole cow's milk, dry infant formula, stainless steel, plastic cutting board, ceramic tile, rubber sheets, and sealed concrete. The system consists of the MicroSEQ Listeria spp. Detection Kit, two sample preparation kits (PrepSEQ Nucleic Acid Extraction Kit and PrepSEQ Rapid Spin Sample Preparation Kit), the Applied Biosystems 7500 Fast Real-Time PCR instrument, and the RapidFinderTM Express v1.1 Software for data analysis. The test method was compared to the ISO 11290-1 reference method using an unpaired study design. The MicroSEQ Listeria spp. Detection Kit and the ISO 11290-1 reference method showed equivalent detection based on Chi-square analysis for all matrixes except hot dogs. For hot dogs, the MicroSEQ method detected more positives than the reference method for the low- and high-level inoculations, with all of the presumptive positives confirmed by the reference method. An independent validation study confirmed these findings on lox and stainless steel surface. The MicroSEQ kit detected all 50 Listeria strains tested and none of the 31 nontarget bacteria strains.
一种用于李斯特菌实时PCR检测的完整系统在五种食品基质和五种环境表面进行了验证，这五种食品基质分别是热狗、烤牛肉、熏鲑鱼、巴氏杀菌全脂牛奶、婴儿配方奶粉，五种环境表面分别是不锈钢、塑料砧板、瓷砖、橡胶板和密封混凝土。该系统由MicroSEQ李斯特菌检测试剂盒、两种样品制备试剂盒（PrepSEQ核酸提取试剂盒和PrepSEQ快速离心样品制备试剂盒）、Applied Biosystems 7500快速实时PCR仪以及用于数据分析的RapidFinderTM Express v1.1软件组成。采用非配对研究设计将该检测方法与ISO 11290-1参考方法进行比较。除热狗外，基于卡方分析，MicroSEQ李斯特菌检测试剂盒和ISO 11290-1参考方法在所有基质中显示出等效的检测结果。对于热狗，在低水平和高水平接种时，MicroSEQ方法检测到的阳性结果比参考方法更多，所有推定阳性结果均经参考方法确认。一项独立的验证研究在熏鲑鱼和不锈钢表面证实了这些发现。MicroSEQ试剂盒检测了所有50株受试李斯特菌菌株，未检测到31株非目标细菌菌株。
