Background The Thermo Scientific™ SureTect™ Salmonella species PCR Assay utilizes Solaris™ reagents for performing PCR for the rapid and specific detection of Salmonella species in a broad range of foods and select environmental surfaces. Objective The aims were to demonstrate the reproducibility of the Thermo Scientific SureTect Salmonella species PCR Assay in a collaborative study using a challenging matrix, cocoa powder, and to extend the scope of the method. Method In the collaborative study, the candidate method was compared to the US Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM) Chapter 5 Salmonella reference method. The candidate method used two PCR thermocyclers, the Applied Biosystems™ QuantStudio™ 5 Real-Time PCR instrument (QS5) and the Applied Biosystems 7500 Fast Real-Time PCR instrument (7500 Fast). Fourteen participants from nine laboratories located within the United States and Europe were solicited for the collaborative study, with 12 participants submitting valid data. Three levels of contamination were evaluated for each matrix. Statistical analysis was conducted according to the probability of detection statistical model. In addition, 11 matrix studies were performed comparing the candidate method to the FDA/BAM Chapter 5 or US Department of Agriculture, Food Safety and Inspection Service, Microbiology Laboratory Guidebook 4.10 Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg, and Siluriformes (Fish) Products and Carcass and Environmental Sponges reference method. Nine of these matrices were also compared to the EN ISO 6579–1:2017/Amd.1:2020(E) Microbiology of the food chain—Horizontal method for the detection, enumeration and serotyping of Salmonella—Part 1: Detection of Salmonella spp.—AMENDMENT 1: Broader range of incubation temperatures, amendment to the status of Annex D, and correction of the composition of MSRV and SC reference method. Results In the collaborative study, the difference in laboratory results indicates equivalence between the candidate method and reference method for the matrix evaluated, and the method demonstrated acceptable interlaboratory reproducibility as determined in the collaborative evaluation. False-positive and false-negative rates were determined for the matrix and produced values of <2%. The two PCR thermocyclers (QS5, 7500 Fast) performed equivalently. There were no result differences between candidate method confirmations and reference method confirmations. In the pre-collaborative matrix extension, the results from the matrix studies showed a comparable performance between the candidate method and the tested reference methods. Conclusions Based on the data generated, the method demonstrated acceptable interlaboratory reproducibility data and statistical analysis. Highlights Due to the COVID-19 pandemic, some participants had to be trained remotely. Additionally, 375 g cocoa powder is known to be a challenging matrix for PCR methods. No unusual cross-contamination or false-positive/negative was reported, highlighting the ease of use, reproducibility, and robustness of the method.

中文翻译：

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Thermo Scientific SureTect 沙门氏菌物种 PCR 检测在多种食品和特定环境表面中的评估：预合作和合作研究：第一行动 2021.02

背景 Thermo Scientific™ SureTect™ 沙门氏菌 PCR 检测利用 Solaris™ 试剂进行 PCR，以快速和特异性地检测各种食品和特定环境表面中的沙门氏菌。目的 目的是证明 Thermo Scientific SureTect 沙门氏菌属 PCR 检测在使用具有挑战性的基质、可可粉的合作研究中的重现性，并扩大该方法的范围。方法 在合作研究中，候选方法与美国食品和药物管理局 (FDA) 细菌学分析手册 (BAM) 第 5 章沙门氏菌参考方法进行了比较。候选方法使用了两个 PCR 热循环仪，Applied Biosystems™ QuantStudio™ 5 实时 PCR 仪 (QS5) 和 Applied Biosystems 7500 快速实时 PCR 仪 (7500 Fast)。来自美国和欧洲的 9 个实验室的 14 名参与者被邀请参加合作研究，其中 12 名参与者提交了有效数据。对每种基质评估了三个污染水平。根据检测概率统计模型进行统计分析。此外，还进行了 11 项矩阵研究，将候选方法与 FDA/BAM 第 5 章或美国农业部、食品安全和检验局微生物实验室指南 4.10 从肉类、家禽、巴氏杀菌蛋和鲶形目中分离和鉴定沙门氏菌进行了比较（鱼）产品和胴体和环境海绵参考方法。其中九个矩阵还与 EN ISO 6579–1:2017/Amd.1 进行了比较：2020(E) 食物链的微生物学——沙门氏菌检测、计数和血清分型的水平方法——第 1 部分：沙门氏菌属的检测——修正 1：更广泛的孵化温度范围，对附件 D 状态的修正和修正MSRV和SC参考方法的组成。结果在合作研究中，实验室结果的差异表明候选方法和参考方法之间的等效性评估的矩阵，并且该方法证明了在合作评估中确定的可接受的实验室间重现性。确定矩阵的假阳性率和假阴性率并产生＜2％的值。两个 PCR 热循环仪 (QS5, 7500 Fast) 表现相同。候选方法确认和参考方法确认之间没有结果差异。在协作前矩阵扩展中，矩阵研究的结果显示候选方法和测试的参考方法之间的性能相当。结论 根据生成的数据，该方法证明了可接受的实验室间重现性数据和统计分析。亮点由于 COVID-19 大流行，一些参与者必须接受远程培训。此外，已知 375 g 可可粉是 PCR 方法具有挑战性的基质。没有报告异常的交叉污染或假阳性/阴性，突出了该方法的易用性、可重复性和稳健性。矩阵研究的结果显示候选方法和测试的参考方法之间的性能相当。结论 根据生成的数据，该方法证明了可接受的实验室间重现性数据和统计分析。亮点由于 COVID-19 大流行，一些参与者必须接受远程培训。此外，已知 375 g 可可粉是 PCR 方法具有挑战性的基质。没有报告异常的交叉污染或假阳性/阴性，突出了该方法的易用性、可重复性和稳健性。矩阵研究的结果显示候选方法和测试的参考方法之间的性能相当。结论 根据生成的数据，该方法证明了可接受的实验室间重现性数据和统计分析。亮点由于 COVID-19 大流行，一些参与者必须接受远程培训。此外，已知 375 g 可可粉是 PCR 方法具有挑战性的基质。没有报告异常的交叉污染或假阳性/阴性，突出了该方法的易用性、可重复性和稳健性。一些参与者必须接受远程培训。此外，已知 375 g 可可粉是 PCR 方法具有挑战性的基质。没有报告异常的交叉污染或假阳性/阴性，突出了该方法的易用性、可重复性和稳健性。一些参与者必须接受远程培训。此外，已知 375 g 可可粉是 PCR 方法具有挑战性的基质。没有报告异常的交叉污染或假阳性/阴性，突出了该方法的易用性、可重复性和稳健性。