验证 Applied Food Diagnostics, Inc. 的 SIMUL-qPCR 李斯特菌属和单核细胞增生李斯特菌同时多重实时聚合酶链反应检测试剂盒在选定食品和环境表面中的应用：AOAC 经性能测试验证的方法标准 062001。

Validation of the Applied Food Diagnostics, Inc. Simultaneous Multiplex Real Time PCR (SIMUL-qPCR) Listeria Species and Monocytogenes Assay in Selected Foods and Environmental Surfaces: AOAC Performance Tested MethodSM 062001.

机构信息

Applied Food Diagnostics, Inc., 18 Industrial Drive, Bloomsburg, PA 17815, USA.

出版信息

J AOAC Int. 2021 Sep 27;104(5):1366-1380. doi: 10.1093/jaoacint/qsab035.

DOI:10.1093/jaoacint/qsab035

PMID:33729515

Abstract

BACKGROUND

The Simultaneous Multiplex Real Time PCR (SIMUL-qPCR) Listeria species and monocytogenes Assay is a quick, reliable method for detecting Listeria species and monocytogenes in environmental and food samples. The assay multiplexes several targets in one run to properly identify Listeria species and monocytogenes. The assay uses proprietary medium, Listeria Recovery and Enrichment Broth (LREB), for enrichment purposes. LREB was specifically formulated to improve the recovery and growth of Listeria while inhibiting competing background flora.

OBJECTIVE

This report details the method validation study to validate frankfurters, ready-to-eat (RTE) sliced turkey, soft fresh raw cheese, chicken salad, ice cream, cooked eggs, pasteurized milk, and frozen/cooked shrimp, as well as environmental surface sponges and swabs for stainless steel, plastic, rubber, ceramic tile, and sealed concrete.

METHOD

Matrix studies, inclusivity/exclusivity, product consistency/stability, and robustness testing were conducted to assess the method's performance.

RESULTS

There were no statistically significant differences found between the candidate and reference methods in the matrix studies. Inclusivity/exclusivity testing showed that the assay was able to detect both Listeria species and monocytogenes strains while excluding the non-Listeria isolates. Small variations in critical test parameters (enrichment time, extraction reagent volume, and extracted sample volume) did not adversely affect the assay's performance, and stability testing indicated consistent results for at least 1 year.

CONCLUSIONS

The data presented in this report show that this a reliable method for detecting Listeria species and monocytogenes.

HIGHLIGHTS

This assay allows for one sample to be tested for both Listeria species and monocytogenes with one PCR test.

摘要

背景

Simultaneous Multiplex Real Time PCR（SIMUL-qPCR）李斯特菌属和单核细胞增生李斯特菌检测试剂盒是一种快速、可靠的方法，用于检测环境和食品样本中的李斯特菌属和单核细胞增生李斯特菌。该检测试剂盒在一次运行中对多个靶标进行多重检测，以正确鉴定李斯特菌属和单核细胞增生李斯特菌。该检测试剂盒使用专有的李斯特菌属恢复和富集肉汤（Listeria Recovery and Enrichment Broth，LREB）进行富集。LREB 是专门配制的，旨在提高李斯特菌的回收率和生长速度，同时抑制竞争背景菌群。

目的

本报告详细介绍了对法兰克福香肠、即食切片火鸡、软质新鲜生奶酪、鸡肉沙拉、冰淇淋、煮鸡蛋、巴氏杀菌奶以及冷冻/熟虾，以及环境表面海绵和不锈钢、塑料、橡胶、瓷砖和密封混凝土的不锈钢拭子进行方法验证研究。

方法

进行了基质研究、包容性/排他性、产品一致性/稳定性和稳健性测试，以评估该方法的性能。

结果

在基质研究中，候选方法和参考方法之间没有发现统计学上的显著差异。包容性/排他性测试表明，该检测试剂盒能够检测到李斯特菌属和单核细胞增生李斯特菌菌株，同时排除非李斯特菌属分离株。关键测试参数（富集时间、提取试剂体积和提取样品体积）的微小变化不会对检测试剂盒的性能产生不利影响，稳定性测试表明，至少在 1 年内结果一致。