3种食品致病菌实时荧光核酸恒温扩增试剂盒方法验证比对研究

摘要

目的 以传统国标培养法作为参考比对方法,考察实时荧光核酸恒温扩增(simultaneous amplification and testing,SAT)食品检测试剂盒,在食品中检测食源性致病菌如志贺氏菌、大肠埃希氏菌O157:H7/NM、阪崎肠杆菌的一致性.方法 以食品中志贺氏菌、大肠埃希氏菌O157:H7/NM和阪崎肠杆菌国标培养法为参考方法,以基因测序法作为确认法,考量SAT食品检测试剂盒方法的灵敏度、特异性、假阳性率、假阴性率、准确度,同时进行2种方法的显著差异检验.结果 3种SAT食品检测试剂盒灵敏度均为100％,假阴性为0;特异性:志贺氏菌100％,阪崎肠杆菌100％,大肠埃希氏菌O157:H7/NM 98.4％.大肠埃希氏菌O157:H7/NM假阳性率为1.2％.结论 SAT试剂盒检测方法具有灵敏度高、特异性强、准确度高的优点,无一例漏检.%Objective To evaluate the consistency of simultaneous amplification and testing (SAT) in detection of foodborne pathogenic microorganisms such as Shigella,Escherichia coli O 157:H7/NM and Enterobacter sakazakii with the traditional national standard culture method as a reference alignment method.Methods The sensitivity,specificity,false positive rate,false negative rate and accuracy of 3 SAT kits for Shigella,Escherichia coli O 157:H7/NM and Enterobacter sakazakii were investigated with the national standards method as reference and the method of molecular sequencing as validation.Meanwhile,the significant difference of positive rate was analyzed by the chi-square test.Results The sensitivities of 3 SAT kits were all 100％,and there were no false negative and misdetection rate.The specificities of Shigella,Escherichia coli O157:H7/NM and Enterobacter sakazakii were 100％,100％,and 98.4％,respectively,which all conformed to the requirements of the test.The false positive rate of Escherichia coli O157:H7/NM was 1.2％.Conclusion The SAT method has the advantages of high sensitivity,specificity,and accuracy,and has no missing detection.