Background The Thermo Scientific SureTect™ Escherichia coli O157:H7 and STEC Screening PCR Assay and SureTect Escherichia coli STEC Identification PCR Assay are real-time PCR kits for the rapid detection of E. coli O157:H7 and non-E. coli O157 Shiga toxin-producing E. coli (STEC) serotypes (O26, O45, O103, O111, O121, O145) from fresh raw spinach, fresh baby leaves, fresh cut tomatoes, frozen raw beef, raw beef trim, and beef carcass sponges. Objective Both assays were evaluated for AOAC® Performance Tested MethodsSM certification. Methods Detection and confirmation inclusivity/exclusivity, matrix, product consistency and stability, and robustness studies were conducted. In the matrix studies, the candidate method was validated against United States and international reference methods for STEC serotypes. Results Matrix studies showed no statistically significant differences between the candidate and reference method results when analyzed by probability of detection. For each inclusivity/exclusivity study, all inclusivity strains and no exclusivity strains were detected by either kit. Robustness testing demonstrated that the identification assay performed reliably despite method deviations; however, although not statistically significant, the screening assay performance was impacted. Product consistency and stability testing demonstrated no statistically significant differences between kit lots and storage time points. Conclusion The data presented show that both assays constitute a rapid and reliable workflow for the detection and confirmation of E. coli O157:H7 and stipulated non-E. coli O157:H7 STEC serotypes from the tested matrixes. Highlights Results are obtained in 80 min post-enrichment with both assays run simultaneously, allowing for the detection and confirmation of STEC within a single workflow.

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Thermo Scientific SureTect™ 大肠杆菌 O157:H7 和 STEC 筛选 PCR 分析和 SureTect™ 大肠杆菌 STEC 鉴定 PCR 分析用于检测大肠杆菌 O157:H7 和大肠杆菌 STEC 血清型（O26、O45、O103、O111、 O121、O145) 来自新鲜生菠菜、新鲜嫩叶、鲜切西红柿、冷冻生牛肉、生牛肉修剪和牛肉胴体海绵：AOAC 性能测试方法 SM 012102

背景 Thermo Scientific SureTect™ 大肠杆菌 O157:H7 和 STEC 筛选 PCR 分析和 SureTect 大肠杆菌 STEC 鉴定 PCR 分析是用于快速检测大肠杆菌 O157:H7 和非大肠杆菌的实时 PCR 试剂盒。大肠杆菌 O157 产志贺毒素大肠杆菌 (STEC) 血清型（O26、O45、O103、O111、O121、O145）来自新鲜生菠菜、新鲜嫩叶、鲜切西红柿、冷冻生牛肉、生牛肉切块和牛肉胴体海绵。目的 两种检测方法都经过了 AOAC® 性能测试方法 SM 认证的评估。方法进行检测和确认包容性/排他性、基质、产品一致性和稳定性以及稳健性研究。在矩阵研究中，候选方法针对 STEC 血清型的美国和国际参考方法进行了验证。结果 矩阵研究显示，在通过检测概率分析时，候选方法和参考方法结果之间没有统计学上的显着差异。对于每个包容性/排他性研究，任一试剂盒均检测到所有包容性菌株和未检测到排他性菌株。稳健性测试表明，尽管存在方法偏差，鉴定分析仍能可靠地执行；然而，虽然没有统计学意义，但筛选分析性能受到了影响。产品一致性和稳定性测试表明试剂盒批次和储存时间点之间没有统计学上的显着差异。结论 所提供的数据表明，这两种检测方法构成了用于检测和确认大肠杆菌 O157:H7 和规定的非大肠杆菌的快速可靠的工作流程。来自测试基质的大肠杆菌 O157:H7 STEC 血清型。