Alles Susan, Curry Stephanie, Almy David, Jagadeesan Balamurugan, Rice Jennifer, Mozola Mark
Neogen Corp., 620 Lesher Pl, Lansing, MI 48912, USA.
J AOAC Int. 2012 Mar-Apr;95(2):424-34. doi: 10.5740/jaoacint.11-262.
A Performance Tested Method validation study was conducted for a new lateral flow immunoassay (Reveal Listeria 2.0) for detection of Listeria spp. in foods and environmental samples. Results of inclusivity testing showed that the test detects all species of Listeria, with the exception of L. grayi. In exclusivity testing conducted under nonselective growth conditions, all non-listeriae tested produced negative Reveal assay results, except for three strains of Lactobacillus spp. However, these lactobacilli are inhibited by the selective Listeria Enrichment Single Step broth enrichment medium used with the Reveal method. Six foods were tested in parallel by the Reveal method and the U.S. Food and Drug Administration/Bacteriological Analytical Manual (FDA/BAM) reference culture procedure. Considering data from both internal and independent laboratory trials, overall sensitivity of the Reveal method relative to that of the FDA/BAM procedure was 101%. Four foods were tested in parallel by the Reveal method and the U.S. Department of Agriculture-Food Safety and Inspection Service (USDA-FSIS) reference culture procedure. Overall sensitivity of the Reveal method relative to that of the USDA-FSIS procedure was 98.2%. There were no statistically significant differences in the number of positives obtained by the Reveal and reference culture procedures in any food trials. In testing of swab or sponge samples from four types of environmental surfaces, sensitivity of Reveal relative to that of the USDA-FSIS reference culture procedure was 127%. For two surface types, differences in the number of positives obtained by the Reveal and reference methods were statistically significant, with more positives by the Reveal method in both cases. Specificity of the Reveal assay was 100%, as there were no unconfirmed positive results obtained in any phase of the testing. Results of ruggedness experiments showed that the Reveal assay is tolerant of modest deviations in test sample volume and device incubation time.
针对一种用于检测食品和环境样本中李斯特菌属的新型侧向流动免疫测定法（Reveal Listeria 2.0）开展了性能验证研究。包容性测试结果表明，该检测方法能检测出除格氏李斯特菌外的所有李斯特菌属菌种。在非选择性生长条件下进行的排他性测试中，除三株乳酸杆菌属菌株外，所有测试的非李斯特菌均产生了Reveal检测阴性结果。然而，这些乳酸杆菌会被与Reveal方法配套使用的选择性李斯特菌单步增菌肉汤增菌培养基抑制。通过Reveal方法和美国食品药品监督管理局/细菌学分析手册（FDA/BAM）参考培养程序对六种食品进行了平行测试。综合内部和独立实验室试验数据，Reveal方法相对于FDA/BAM程序的总体灵敏度为101%。通过Reveal方法和美国农业部食品安全检验局（USDA-FSIS）参考培养程序对四种食品进行了平行测试。Reveal方法相对于USDA-FSIS程序的总体灵敏度为98.2%。在任何食品试验中，Reveal方法和参考培养程序获得的阳性数量均无统计学显著差异。在对四种环境表面的拭子或海绵样本进行检测时，Reveal方法相对于USDA-FSIS参考培养程序的灵敏度为127%。对于两种表面类型，Reveal方法和参考方法获得的阳性数量差异具有统计学意义，两种情况下Reveal方法获得的阳性更多。Reveal检测的特异性为100%，因为在测试的任何阶段均未获得未经确认的阳性结果。耐用性实验结果表明，Reveal检测对测试样本体积和设备孵育时间的适度偏差具有耐受性。
