赛默飞世尔科技SureTect李斯特菌属检测方法的评估。AOAC性能验证方法071304。

Evaluation of the Thermo Scientific SureTect Listeria species assay. AOAC Performance Tested Method 071304.

作者信息

Cloke Jonathan, Evans Katharine, Crabtree David, Hughes Annette, Simpson Helen, Holopainen Jani, Wickstrand Nina, Kauppinen Mikko, Leon-Velarde Carlos, Larson Nathan, Dave Keron

出版信息

J AOAC Int. 2014 Mar-Apr;97(2):521-38.

PMID:24830165

Abstract

The Thermo Scientific SureTect Listeria species Assay is a new real-time PCR assay for the detection of all species of Listeria in food and environmental samples. This validation study was conducted using the AOAC Research Institute (RI) Performance Tested Methods program to validate the SureTect Listeria species Assay in comparison to the reference method detailed in International Organization for Standardization 11290-1:1996 including amendment 1:2004 in a variety of foods plus plastic and stainless steel. The food matrixes validated were smoked salmon, processed cheese, fresh bagged spinach, cantaloupe, cooked prawns, cooked sliced turkey meat, cooked sliced ham, salami, pork frankfurters, and raw ground beef. All matrixes were tested by Thermo Fisher Scientific, Microbiology Division, Basingstoke, UK. In addition, three matrixes (pork frankfurters, fresh bagged spinach, and stainless steel surface samples) were analyzed independently as part of the AOAC-RI-controlled independent laboratory study by the University ofGuelph, Canada. Using probability of detection statistical analysis, a significant difference in favour of the SureTect assay was demonstrated between the SureTect and reference method for high level spiked samples of pork frankfurters, smoked salmon, cooked prawns, stainless steel, and low-spiked samples of salami. For all other matrixes, no significant difference was seen between the two methods during the study. Inclusivity testing was conducted with 68 different isolates of Listeria species, all of which were detected by the SureTect Listeria species Assay. None of the 33 exclusivity isolates were detected by the SureTect Listeria species Assay. Ruggedness testing was conducted to evaluate the performance of the assay with specific method deviations outside of the recommended parameters open to variation, which demonstrated that the assay gave reliable performance. Accelerated stability testing was additionally conducted, validating the assay shelf life.

摘要

赛默飞世尔科技SureTect李斯特菌属检测试剂盒是一种新型实时荧光定量PCR检测试剂盒，用于检测食品和环境样本中的所有李斯特菌属菌种。本验证研究采用美国官方分析化学师协会（AOAC）研究机构（RI）的性能验证方法程序，将SureTect李斯特菌属检测试剂盒与国际标准化组织11290-1:1996（包括2004年第1号修正案）中详述的参考方法进行比较，以验证该试剂盒在多种食品以及塑料和不锈钢样本中的性能。经验证的食品基质包括烟熏三文鱼、加工奶酪、新鲜袋装菠菜、哈密瓜、熟虾、熟火鸡肉片、熟火腿片、意大利腊肠、猪肉法兰克福香肠和生牛肉末。所有基质样本均由英国贝辛斯托克的赛默飞世尔科技微生物学部进行检测。此外，加拿大圭尔夫大学作为AOAC-RI控制的独立实验室研究的一部分，对三种基质样本（猪肉法兰克福香肠、新鲜袋装菠菜和不锈钢表面样本）进行了独立分析。通过检测概率统计分析表明，对于猪肉法兰克福香肠、烟熏三文鱼、熟虾、不锈钢的高加标水平样本以及意大利腊肠的低加标水平样本，SureTect检测试剂盒与参考方法相比，在检测结果上存在显著差异且更具优势。在本研究中，对于所有其他基质样本，两种方法之间未观察到显著差异。采用68种不同的李斯特菌属分离株进行了包容性测试，所有分离株均能被SureTect李斯特菌属检测试剂盒检测到。33种排他性分离株中，SureTect李斯特菌属检测试剂盒均未检测到。进行了耐用性测试，以评估该检测试剂盒在推荐参数范围外特定方法偏差情况下的性能，结果表明该检测试剂盒性能可靠。此外，还进行了加速稳定性测试，以验证该检测试剂盒的保质期。