Bastin Benjamin, Horine Leo, Bird Patrick, Benzinger M Joseph, Agin James, Goins David, Smith Paul, Centola Michael
Q Laboratories, Inc., 1400 Harrison Ave, Cincinnati, OH 45214.
PolySkope Labs, 755 Research Parkway, Oklahoma City, OK 73104.
J AOAC Int. 2019 Sep 1;102(5):1455-1471. doi: 10.1093/jaoac/102.5.1455.
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.
A Performance Tested MethodSM study was conducted to validate Polyskope 1.0 for inclusivity and exclusivity as well as a matrix comparison study.
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 × 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.
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.
Polyskope 1.0多重检测法是一种新型检测方法，可通过实时聚合酶链反应在一次富集培养中同时检测大肠杆菌O157、非O157产志贺毒素大肠杆菌（STEC）、单核细胞增生李斯特菌和沙门氏菌属。
开展一项性能验证方法研究，以验证Polyskope 1.0的包容性、排他性以及进行基质比较研究。
根据美国官方分析化学师协会（AOAC INTERNATIONAL）的验证指南，在一项未配对的独立验证研究中，将该检测法与参考方法进行比较，对Polyskope 1.0进行评估。在用三种目标微生物的悬浮液接种后，Polyskope 1.0对生碎牛肉（25克）、熟火鸡肉（25克）、嫩菠菜（25克）和不锈钢环境表面海绵（4×4英寸测试区域）进行了评估。所有基质均与美国食品药品监督管理局《细菌学分析手册》、美国农业部食品安全与检验局《微生物学实验室指南手册》或国际标准化组织标准中的适当参考方法进行比较。
对于三种食品基质和一个环境表面，Polyskope 1.0在候选方法和参考方法结果之间，以及推定结果和确认结果之间均未显示出统计学上的显著差异。包容性和排他性评估结果表明，该检测方法能够准确检测目标分析物，并排除所有非目标生物。在稳定性或批次间评估中未观察到差异。Polyskope 1.0通过不受方法参数的微小变化影响而表现出稳健性，所有八种变化对结果均无统计学上的显著影响。结论与要点：Polyskope 1.0被证明是一种用于在四种基质中检测单核细胞增生李斯特菌、沙门氏菌属、非O157 STEC和大肠杆菌O157的特异性强、高度准确且稳健的方法。
