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首页> 外文期刊>Journal of AOAC International >Independent Validation for the Polyskope 1.0 Multiplex Pathogen Detection Assay for the Detection of Shiga Toxin-Producing Escherichia coli Non-O157 STEC, Escherichia coli O157, Listeria monocytogenes, and Salmonella Species
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Independent Validation for the Polyskope 1.0 Multiplex Pathogen Detection Assay for the Detection of Shiga Toxin-Producing Escherichia coli Non-O157 STEC, Escherichia coli O157, Listeria monocytogenes, and Salmonella Species

机译：用于检测滋生毒素的大肠杆菌非O157 STEC，大肠杆菌O157，Histeria单核细胞增生和沙门氏菌种类的多重验证的多重验证

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Background: The Polyskope 1.0 Multiplex Assay is a novel test to simultaneously detect Escherichia coli O157, non-O157 Shiga Toxin-Producing E. coli (STEC), Listeria monocytogenes, and Salmonella species in a single enrichment using real-time PCR. Objective: A Performance Tested Method(SM) study was conducted to validate Polyskope 1.0 for inclusivity and exclusivity as well as a matrix comparison study. Method: This assay was evaluated in an unpaired independent validation study compared with reference methods according to AOAC INTERNATIONAL validation guidelines. Polyskope 1.0 evaluated raw ground beef (25 g), deli turkey (25 g), baby spinach (25 g), and stainless-steel environmental surface sponges (4 x 4 in. test area) after inoculation with a suspension of the three target microorganisms. All matrices were compared with appropriate reference methods from the U.S. Food and Drug Administration Bacteriological Analytical Manual, U.S. Department of Agriculture Food Safety and Inspection Service Microbiology Laboratory Guidebook, or International Organization for Standardization standards. Results: Polyskope 1.0 demonstrated no statistically significant differences between candidate and reference method results or between presumptive and confirmed results for three food matrices and one environmental surface. Results from inclusivity and exclusivity evaluations indicated the test method can accurately detect the target analytes and excluded all nontarget organisms. No differences were observed with the stability or lot-to-lot evaluations. Polyskope 1.0 demonstrated robustness by remaining unaffected by small variations in method parameters, which had no statistically significant effect on the results for all eight variations. Conclusions and Highlights: Polyskope 1.0 was shown to be a specific, highly accurate, and robust method for the detection of Listeria monocytogenes, Salmonella species, non-O157 STECs, and E. coli O157 across four matrices.
机译：背景技术多重测定是一种新的试验，用于使用实时PCR在单一富集中检测大肠杆菌O157，非O157滋菌毒素的大肠杆菌（STEC），李斯特菌单核细胞增生和沙门氏菌。目的：进行性能测试方法（SM）研究以验证多孔1.0，以含有含量和排他性以及基质比较研究。方法：根据AOAC国际验证指南，与参考方法相比，在未配对的独立验证研究中评估了该测定。 Polyskope 1.0评估了原料牛肉（25g），熟食火鸡（25g），婴儿菠菜（25g）和不锈钢环境表面海绵（4×4英寸试验区域），在接种三个目标后接种后微生物。将所有基质与来自美国食品和药物管理局的适当参考方法进行比较，美国农业食品安全和检测服务微生物实验室指南或国际标准化标准组织。结果：Polyskope 1.0证明了候选和参考方法结果之间没有统计学显着的差异，或者在三种食物基质和一个环境表面的推定和确认结果之间。包括夹杂度和排他性评估结果表明，测试方法可以准确地检测目标分析物并排除所有Nontarget生物体。稳定性或批量评估没有观察到差异。 Polyskope 1.0通过剩下的方法参数的小变化仍然不受影响来展示鲁棒性，这对所有八种变化的结果没有统计学显着的影响。结论和亮点：PolySkope 1.0被证明是检测鼠李生素单核细胞增生，沙门氏菌物种，非O157 STEC和四种基质的大肠杆菌O157的特定，高度准确和稳健的方法。

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