Cloke Jonathan, Leon-Velarde Carlos, Larson Nathan, Dave Keron, Evans Katharine, Crabtree David, Hughes Annette, Hopper Craig, Simpson Helen, Withey Sophie, Oleksiuk Milena, Holopainen Jani, Wickstrand Nina, Kauppinen Mikko
J AOAC Int. 2014 Jan-Feb;97(1):133-54. doi: 10.5740/jaoacint.13-246.
The Thermo Scientific SureTect Listeria monocytogenes Assay is a new real-time PCR assay for the detection of Listeria monocytogenes in food and environmental samples. This assay was validated using the AOAC Research Institute (AOAC-RI) Performance Tested Methods program in comparison to the reference method detailed in International Organization for Standardization 11290-1:1996, including Amendment 1:2004 with the following foods and food contact surfaces: smoked salmon, processed cheese, fresh bagged spinach, fresh cantaloupe, cooked prawns (chilled product), cooked sliced turkey meat (chilled product), ice cream, pork frankfurters, salami, ground raw beef meat (12% fat), plastic, and stainless steel. All matrixes were tested by Thermo Fisher Scientific, Microbiology Division, Basingstoke, UK. In addition, three matrixes (pork frankfurters, bagged lettuce, and stainless steel) were analyzed independently as part of the AOAC-RI controlled laboratory study by the University of Guelph, Canada. Using probability of detection (POD) statistical analysis, a significant difference was demonstrated between the candidate and reference methods for salami, cooked sliced turkey and ice cream in favor of the SureTect assay. For all other matrixes, no significant difference by POD was seen between the two methods during the study. Inclusivity and exclusivity testing was also conducted with 53 and 30 isolates, respectively, which demonstrated that the SureTect assay was able to detect all serotypes of L. monocytogenes. None of the exclusivity isolates analyzed were detected by the SureTect assay. Ruggedness testing was conducted to evaluate the performance of the assay with specific method deviations outside the recommended parameters open to variation, i.e., enrichment time and temperature and lysis temperature, which demonstrated that the assay gave reliable performance. Accelerated stability testing was also conducted, validating the assay shelf life.
赛默飞世尔科技的SureTect单核细胞增生李斯特菌检测方法是一种用于检测食品和环境样本中单核细胞增生李斯特菌的新型实时荧光定量PCR检测方法。该检测方法通过美国官方分析化学家协会（AOAC）研究机构（AOAC-RI）性能验证方法计划进行验证，并与国际标准化组织11290-1:1996（包括修订版1:2004）中详述的参考方法进行比较，所涉及的食品和食品接触表面包括：烟熏三文鱼、加工奶酪、新鲜袋装菠菜、新鲜哈密瓜、熟虾（冷藏产品）、熟火鸡肉片（冷藏产品）、冰淇淋、猪肉法兰克福香肠、意大利腊肠、含12%脂肪的生牛肉末、塑料和不锈钢。所有基质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，加拿大圭尔夫大学独立分析了三种基质（猪肉法兰克福香肠、袋装生菜和不锈钢），作为AOAC-RI对照实验室研究的一部分。通过检测概率（POD）统计分析表明，在意大利腊肠、熟火鸡肉片和冰淇淋的检测中，候选方法与参考方法之间存在显著差异，SureTect检测方法更具优势。在研究过程中，对于所有其他基质，两种方法的POD无显著差异。还分别对53株和30株菌株进行了包容性和排他性测试，结果表明SureTect检测方法能够检测出所有血清型的单核细胞增生李斯特菌。SureTect检测方法未检测出任何分析的排他性菌株。进行了耐用性测试，以评估该检测方法在推荐参数范围之外的特定方法偏差（即增菌时间和温度以及裂解温度）下的性能，结果表明该检测方法性能可靠。还进行了加速稳定性测试，验证了该检测方法的保质期。
