Background The Clear Safety Salmonella method was modified to improve sample preparation, PCR reagents, library preparation, flow cell quality control, library loading mix, priming mix, and sequencing kit reagents and steps. Objective To evaluate the modified Clear Safety Salmonella method (manual and automated) via independent and method developer validation studies according to current AOAC INTERNATIONAL Validation Guidelines. Method Performance of the modified Clear Safety Salmonella method (manual and automated) was assessed for selectivity (using 105 inclusive and 30 exclusive strains), probability of detection in matrixes, product consistency, stability, and robustness. The modified Clear Safety Salmonella method was compared with the appropriate reference method for Salmonella detection on 4 inch × 4 inch stainless steel environmental surfaces, and in chicken carcass rinse (30 mL), raw ground chicken (375 g), dry pet food (375 g), and ready-to-eat deli turkey breast (375 g). Results The modified Clear Safety Salmonella method (manual and automated) demonstrated no statistically significant differences between the candidate and reference method probability of detection or between the presumptive and confirmed results for all target food matrixes and the stainless steel surface. Additionally, the modified method (manual and automated) detected all 105 inclusivity organisms and excluded all 30 exclusivity organisms. The product consistency and kit stability studies showed no statistical differences between lots or over the term of the kit’s shelf life. In robustness studies, changes in enrichment time, diluted sample volume, and sample volume for PCR did not show any statistical difference in terms of assay performance. Conclusions The modified Clear Safety Salmonella method (both manual and automated) is statistically equivalent to or better than the reference methods. Highlights The Clear Safety Salmonella method utilizes PCR amplification and targeted next-generation sequencing technology to selectively detect Salmonella enterica.

中文翻译：

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验证改良的透明安全沙门氏菌用于检测选定的家禽和宠物食品基质以及不锈钢上的肠道沙门氏菌：AOAC 性能测试方法 SM 111802

背景 对 Clear Safety 沙门氏菌方法进行了修改，以改进样品制备、PCR 试剂、文库制备、流通池质量控制、文库加载混合物、引发混合物以及测序试剂盒试剂和步骤。目的 根据当前的 AOAC 国际验证指南，通过独立和方法开发人员验证研究评估改进的清除安全沙门氏菌方法（手动和自动）。方法 对改良的 Clear Safety 沙门氏菌方法（手动和自动）的性能进行了评估，包括选择性（使用 105 种和 30 种专用菌株）、基质中的检测概率、产品一致性、稳定性和稳健性。在 4 英寸 × 4 英寸不锈钢环境表面和鸡胴体冲洗液 (30 mL)、生碎鸡肉 (375 g)、干宠物食品 (375 g) 和即食熟食火鸡胸肉 (375 g)。结果 改进的透明安全沙门氏菌方法（手动和自动）显示候选方法和参考方法的检测概率之间或所有目标食品基质和不锈钢表面的推定结果和确认结果之间没有统计学上的显着差异。此外，修改后的方法（手动和自动）检测了所有 105 个包容性生物并排除了所有 30 个排他性生物。产品一致性和试剂盒稳定性研究表明批次之间或试剂盒保质期没有统计学差异。在稳健性研究中，富集时间、稀释样品体积和 PCR 样品体积的变化在测定性能方面没有显示出任何统计学差异。结论 改良的清除安全沙门氏菌方法（手动和自动）在统计学上等同于或优于参考方法。亮点 Clear Safety Salmonella 方法利用 PCR 扩增和靶向下一代测序技术选择性地检测肠沙门氏菌。结论 改良的清除安全沙门氏菌方法（手动和自动）在统计学上等同于或优于参考方法。亮点 Clear Safety Salmonella 方法利用 PCR 扩增和靶向下一代测序技术选择性地检测肠沙门氏菌。结论 改良的清除安全沙门氏菌方法（手动和自动）在统计学上等同于或优于参考方法。亮点 Clear Safety Salmonella 方法利用 PCR 扩增和靶向下一代测序技术选择性地检测肠沙门氏菌。