基于实时荧光核酸恒温扩增技术的食源性致病微生物检测

目的考察实时荧光核酸恒温扩增(simultaneous amplification and testing,SAT)技术在食源性致病微生物检测中的应用价值。方法以国标培养法及分子测序为参考方法,研究SAT方法用于检测沙门氏菌、金黄色葡萄球菌、副溶血性弧菌及单增李斯特氏菌的灵敏度、特异性、假阳性率、假阴性率、准确度,同时利用卡方检验分析阳性率显著性差异。结果 4种致病菌SAT试剂盒的灵敏度均为100%,不存在假阴性及漏检情况;沙门氏菌(94.2%)、金黄色葡萄球菌(99.5%)、副溶血性弧菌(100%)和单增李斯特氏菌(100%)的特异性符合要求,假阳性率均小于9.6%,准确度均大于94%。结论 SAT技术具有高灵敏度、特异性强、准确度高,可作为上述4种致病菌的检验方法。

Simultaneous amplification and testing application in detection of foodborne pathogenic microorganisms

Objective To evaluate the application value of simultaneous amplification and testing (SAT) in detection of foodborne pathogenic microorganisms. Methods The sensitivity, specificity, false positive rate, false negative rate and accuracy of Salmonella spp., Staphylococcus aureus, Vibrio parahaemolyticus and Listeria monocytogenes were studied by using the methods of national standards and molecular sequencing. Meanwhile, the significant difference of positive rate was analyzed by the chi-square. Results The sensitivity of four pathogens were all 100%, and there were no false negative and misdetection rate. The specificity of Salmonella spp., Staphylococcus aureus, Vibrio parahaemolyticus and Listeria monocytogenes were 94.2%, 99.5%, 100%, and 100% respectively, and they all conformed to the requirements of the test. The false positive rates were all below 9.6% and the accuracy rates were superior to the standard (94%). Conclusion It is feasible to detect the four pathogens above with SAT for its high sensitivity, specificity and accuracy.