Wall Jason, Conrad Rick, Latham Kathy, Liu Eric
Life Technologies, 2130 Woodward St, Austin, TX 78744, USA.
J AOAC Int. 2014 Mar 1;97(2):484-491. doi: 10.5740/jaoacint.13-251.
Real-time PCR methods for detecting foodborne pathogens offer the advantages of simplicity and quick time to results compared to traditional culture methods. The addition of a recirculating pooled immunomagnetic separation method prior to real-time PCR analysis increases processing output while reducing both cost and labor. This AOAC Research Institute method modification study validates the MicroSEQ® Salmonella spp. Detection Kit [AOAC Performance Tested Method (PTM) 031001] linked with the Pathatrix® 10-Pooling Salmonella spp. Kit (AOAC PTM 090203C) in diced tomatoes, chocolate, and deli ham. The Pathatrix 10-Pooling protocol represents a method modification of the enrichment portion of the MicroSEQ Salmonella spp.
The results of the method modification were compared to standard cultural reference methods for diced tomatoes, chocolate, and deli ham. All three matrixes were analyzed in a paired study design. An additional set of chocolate test portions was analyzed using an alternative enrichment medium in an unpaired study design. For all matrixes tested, there were no statistically significant differences in the number of positive test portions detected by the modified candidate method compared to the appropriate reference method. The MicroSEQ Salmonella spp. protocol linked with the Pathatrix individual or 10-Pooling procedure demonstrated reliability as a rapid, simplified, method for the preparation of samples and subsequent detection of Salmonella in diced tomatoes, chocolate, and deli ham.
与传统培养方法相比，用于检测食源性病原体的实时聚合酶链反应（PCR）方法具有操作简单、出结果快的优点。在实时PCR分析之前添加循环混合免疫磁珠分离方法，可提高处理效率，同时降低成本和劳动力。美国官方分析化学师协会（AOAC）研究机构的这项方法改进研究，验证了与Pathatrix® 10混合沙门氏菌试剂盒（AOAC性能测试方法（PTM）090203C）联用的MicroSEQ®沙门氏菌检测试剂盒（AOAC PTM 031001）在切块番茄、巧克力和熟火腿中的效果。Pathatrix 10混合方案是对MicroSEQ沙门氏菌富集部分方法的改进。
将方法改进的结果与切块番茄、巧克力和熟火腿的标准培养参考方法进行比较。所有三种基质均采用配对研究设计进行分析。在非配对研究设计中，使用替代富集培养基对另一组巧克力测试部分进行分析。对于所有测试的基质，与适当的参考方法相比，改进后的候选方法检测到的阳性测试部分数量没有统计学上的显著差异。与Pathatrix单独或10混合程序联用的MicroSEQ沙门氏菌方案，作为一种快速、简化的方法，用于切块番茄、巧克力和熟火腿中样品的制备及后续沙门氏菌的检测，证明是可靠的。
