Fach P, Dilasser F, Grout J, Tache J
Centre National d'Etudes Vétérinaires et Alimentaires, Laboratoire Central d'Hygiène Alimentaire, Unité Atelier de Biotechnologie, Paris, France.
J Food Prot. 1999 Dec;62(12):1387-93. doi: 10.4315/0362-028x-62.12.1387.
A commercially available polymerase chain reaction (PCR) kit was evaluated for the detection of Salmonella spp. in food samples. The test combines PCR amplification and sandwich hybridization of the amplified DNA in microtiter plates. The sensitivity and specificity was evaluated with 52 Salmonella strains and 51 non-Salmonella strains and showed that the test was entirely reliable. The threshold sensitivity was 10(2) CFU/ml. The limit of detection of dead cells that determines the minimum detection level of dead cells in food samples was superior to 10(6) CFU/25 g, a level rarely achieved in naturally contaminated samples. After an 18-h pre-enrichment step, the test could detect viable Salmonella in artificially contaminated food samples, even for the lower contamination level (3 CFU/25 g). There was complete agreement between the PCR test and the ISO 6579 bacteriological reference method with artificially contaminated samples. Regarding the accuracy of the results obtained from 253 naturally or noncontaminated foods and from 32 artificially contaminated foods, the agreement percentage was 99.6%. The fidelity of the technique was evaluated in a collaborative study with eight European laboratories and showed a correlation of 98.4%.
对一种市售聚合酶链反应(PCR)试剂盒进行了评估,以检测食品样本中的沙门氏菌属。该检测方法将PCR扩增与微量滴定板中扩增DNA的夹心杂交相结合。用52株沙门氏菌菌株和51株非沙门氏菌菌株评估了其敏感性和特异性,结果表明该检测方法完全可靠。阈值敏感性为10(2) CFU/ml。确定食品样本中死细胞最低检测水平的死细胞检测限优于10(6) CFU/25 g,这一水平在自然污染样本中很少能达到。经过18小时的预富集步骤后,该检测方法能够检测人工污染食品样本中的活菌沙门氏菌,即使是较低的污染水平(3 CFU/25 g)。对于人工污染样本,PCR检测与ISO 6579细菌学参考方法完全一致。对于从253份天然或未污染食品以及32份人工污染食品中获得的结果的准确性,一致率为99.6%。在与八个欧洲实验室的合作研究中评估了该技术的保真度,显示相关性为98.4%。