Cloke Jonathan, Clark Dorn, Radcliff Roy, Leon-Velarde Carlos, Larson Nathan, Dave Keron, Evans Katharine, Crabtree David, Hughes Annette, Simpson Helen, Holopainen Jani, Wickstrand Nina, Kauppinen Mikko
J AOAC Int. 2014 Mar-Apr;97(2):539-60.
The Thermo Scientific SureTect Salmonella species Assay is a new real-time PCR assay for the detection of Salmonellae in food and environmental samples. This validation study was conducted using the AOAC Research Institute (RI) Performance Tested Methods program to validate the SureTect Salmonella species Assay in comparison to the reference method detailed in International Organization for Standardization 6579:2002 in a variety of food matrixes, namely, raw ground beef, raw chicken breast, raw ground pork, fresh bagged lettuce, pork frankfurters, nonfat dried milk powder, cooked peeled shrimp, pasteurized liquid whole egg, ready-to-eat meal containing beef, and stainless steel surface samples. With the exception of liquid whole egg and fresh bagged lettuce, which were tested in-house, all matrixes were tested by Marshfield Food Safety, Marshfield, WI, on behalf of Thermo Fisher Scientific. In addition, three matrixes (pork frankfurters, lettuce, and stainless steel surface samples) were analyzed independently as part of the AOAC-RI-controlled laboratory study by the University of Guelph, Canada. No significant difference by probability of detection or McNemars Chi-squared statistical analysis was found between the candidate or reference methods for any of the food matrixes or environmental surface samples tested during the validation study. Inclusivity and exclusivity testing was conducted with 117 and 36 isolates, respectively, which demonstrated that the SureTect Salmonella species Assay was able to detect all the major groups of Salmonella enterica subspecies enterica (e.g., Typhimurium) and the less common subspecies of S. enterica (e.g., arizoniae) and the rarely encountered S. bongori. None of the exclusivity isolates analyzed were detected by the SureTect Salmonella species Assay. Ruggedness testing was conducted to evaluate the performance of the assay with specific method deviations outside of the recommended parameters open to variation (enrichment time and temperature, and lysis temperature), which demonstrated that the assay gave reliable performance. Accelerated stability testing was additionally conducted, validating the assay shelf life.
赛默飞世尔科技沙门氏菌属检测试剂盒是一种用于检测食品和环境样本中沙门氏菌的新型实时荧光定量PCR检测试剂盒。本验证研究采用美国官方分析化学师协会（AOAC）研究机构（RI）的性能验证方法程序，将赛默飞世尔科技沙门氏菌属检测试剂盒与国际标准化组织6579:2002中详述的参考方法进行比较，在多种食品基质中进行验证，这些基质包括生牛肉末、生鸡胸肉、生猪肉末、新鲜袋装生菜、猪肉肠、脱脂奶粉、熟去壳虾、巴氏杀菌全蛋液、含牛肉的即食餐以及不锈钢表面样本。除了全蛋液和新鲜袋装生菜在内部进行检测外，所有基质均由威斯康星州马什菲尔德市的马什菲尔德食品安全公司代表赛默飞世尔科技进行检测。此外，加拿大圭尔夫大学作为AOAC-RI控制的实验室研究的一部分，独立分析了三种基质（猪肉肠、生菜和不锈钢表面样本）。在验证研究期间，对于所测试的任何食品基质或环境表面样本，候选方法和参考方法在检测概率或McNemar卡方统计分析方面均未发现显著差异。分别使用117株和36株菌株进行了包容性和排他性测试，结果表明赛默飞世尔科技沙门氏菌属检测试剂盒能够检测肠炎沙门氏菌肠道亚种的所有主要菌群（如鼠伤寒沙门氏菌）、肠炎沙门氏菌较不常见的亚种（如亚利桑那沙门氏菌）以及罕见的邦戈尔沙门氏菌。赛默飞世尔科技沙门氏菌属检测试剂盒未检测到任何分析的排他性菌株。进行了耐用性测试，以评估该检测试剂盒在推荐参数之外的特定方法偏差（增菌时间和温度以及裂解温度）下的性能，结果表明该检测试剂盒性能可靠。此外还进行了加速稳定性测试，验证了该检测试剂盒的保质期。
