Alles Susan, Mozola Mark
Neogen Corp., 620 Lesher Pl, Lansing, MI 48912, USA.
J AOAC Int. 2009 Nov-Dec;92(6):1840-5.
A study was conducted to validate the GeneQuence Salmonella DNA hybridization assay, Performance Tested Method 030201, for detection of Salmonella spp. in peanut butter. The study was organized by the AOAC Research Institute under its Emergency Response Validation program. Peanut butter samples inoculated with S. Typhimurium were prepared by an independent laboratory and shipped to study participants for testing. The set of blind-coded test samples consisted of five uninoculated controls, 20 portions inoculated with S. Typhimurium at a low level [determined by most probable number (MPN) analysis to contain 1.1 CFU/25 g portion], and 20 portions inoculated with S. Typhimurium at a higher level (11 CFU/25 g portion by MPN analysis). Samples were tested in parallel by the GeneQuence method and by the U.S. Food and Drug Administration's Bacteriological Analytical Manual reference culture procedure. All five control samples were negative by both methods. For the low-level samples, the same two samples were positive by both the GeneQuence and reference methods. For the high-level samples, the same 19 samples were positive by both methods. All positive GeneQuence assays were confirmed by plating from associated broth cultures. Agreement between the GeneQuence and reference methods was 100%. Sensitivity and specificity of the GeneQuence method were both 100%. Because neither the low- nor the high-level samples yielded the desired fractional positive results (5-15 positives out of 20 samples), a second trial was conducted. Samples in the second trial contained 0.1 and 0.5 CFU/25 g portion for the low and high levels, respectively. All five control samples were negative by both methods. For the low-level samples, the same two samples were positive by both the GeneQuence and reference methods. For the high-level samples, the same three samples were positive by both methods. All positive GeneQuence assays were confirmed by plating from associated broth cultures. Sensitivity and specificity of the GeneQuence method were both 100%. Although once again the desired level of fractional positive results was not obtained, there was 100% agreement between the GeneQuence and reference methods. Based on the results of both trials, it is recommended that the validated claims for Performance Tested Method 030201 be expanded to include peanut butter.
开展了一项研究，以验证基因序列沙门氏菌DNA杂交检测法（性能验证方法030201）用于检测花生酱中沙门氏菌属的效果。该研究由美国官方分析化学师协会研究所在其应急响应验证项目下组织开展。接种了鼠伤寒沙门氏菌的花生酱样本由一家独立实验室制备，并运送给研究参与者进行检测。这组盲编码测试样本包括5个未接种的对照样本、20份接种低水平鼠伤寒沙门氏菌的样本（通过最大可能数分析确定每25克样本含1.1 CFU），以及20份接种高水平鼠伤寒沙门氏菌的样本（通过最大可能数分析为每25克样本含11 CFU）。样本通过基因序列法和美国食品药品监督管理局的《细菌学分析手册》参考培养程序进行平行检测。两种方法检测的所有5个对照样本均为阴性。对于低水平样本，基因序列法和参考方法检测的相同两个样本均为阳性。对于高水平样本，两种方法检测的相同19个样本均为阳性。所有基因序列法检测呈阳性的结果均通过相关肉汤培养物的平板接种得到确认。基因序列法与参考方法的一致性为100%。基因序列法的灵敏度和特异性均为100%。由于低水平和高水平样本均未产生预期的部分阳性结果（20个样本中有5 - 15个阳性），因此进行了第二次试验。第二次试验中的样本低水平和高水平分别含每25克样本0.1和0.5 CFU。两种方法检测的所有5个对照样本均为阴性。对于低水平样本，基因序列法和参考方法检测的相同两个样本均为阳性。对于高水平样本，两种方法检测的相同3个样本均为阳性。所有基因序列法检测呈阳性的结果均通过相关肉汤培养物的平板接种得到确认。基因序列法的灵敏度和特异性均为100%。尽管再次未获得预期的部分阳性结果水平，但基因序列法与参考方法之间的一致性仍为100%。基于两项试验的结果，建议将性能验证方法030201的有效声明扩展至包括花生酱。
