Clear Labs, 1559 Industrial Rd, San Carlos, CA 94070, USA.
Q Laboratories, 1930 Radcliff Drive, Cincinnati, OH 45204, USA.
J AOAC Int. 2022 Feb 4;105(1):230-248. doi: 10.1093/jaoacint/qsab062.
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.
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.
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).
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.
The modified Clear Safety Salmonella method (both manual and automated) is statistically equivalent to or better than the reference methods.
The Clear Safety Salmonella method utilizes PCR amplification and targeted next-generation sequencing technology to selectively detect Salmonella enterica.
对 Clear Safety Salmonella 方法进行了修改，以改进样品制备、PCR 试剂、文库制备、流池质量控制、文库加载混合液、引物混合液和测序试剂盒试剂和步骤。
根据当前的 AOAC INTERNATIONAL 验证指南，通过独立和方法开发人员验证研究来评估改良的 Clear Safety Salmonella 方法（手动和自动）。
通过选择性（使用 105 种包容性和 30 种排他性菌株）、基质中检测概率、产品一致性、稳定性和稳健性评估改良的 Clear Safety Salmonella 方法（手动和自动）的性能。改良的 Clear Safety Salmonella 方法与适当的参考方法进行比较，用于检测 4 英寸×4 英寸不锈钢环境表面上的沙门氏菌，以及鸡胴体冲洗液（30 mL）、生鸡肉（375 g）、干宠物食品（375 g）和即食熟火鸡胸肉（375 g）。
改良的 Clear Safety Salmonella 方法（手动和自动）在候选方法和参考方法的检测概率之间，以及所有目标食品基质和不锈钢表面的推定和确认结果之间，均无统计学差异。此外，改良方法（手动和自动）检测到所有 105 种包容性生物体，排除了所有 30 种排他性生物体。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内均无统计学差异。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统计学差异。
改良的 Clear Safety Salmonella 方法（手动和自动）与参考方法在统计学上等效或优于参考方法。
Clear Safety Salmonella 方法利用 PCR 扩增和靶向下一代测序技术，选择性检测沙门氏菌。
