食品中单增李斯特氏菌检测PCR-免疫胶体金试纸条方法的建立

PCR-immunogold method for detection of Listeria monocytogenes in food

目的建立食品中单核增生李斯特氏菌快速检测PCR-免疫胶体金试纸条法。方法通过设计特异性引物建立单增李斯特氏菌检测PCR方法并使用免疫胶体金技术建立PCR产物快速检测试纸条;用试验菌株检测PCR-免疫胶体金试纸条方法的检测特异度与敏感度。使用新建方法对市售肉制品和乳制品中单核增生李斯特氏菌进行检测,验证该方法在食品检测中的可行性。结果 PCR-免疫胶体金法具有良好的特异度,敏感度比标准琼脂糖凝胶电泳法高100倍。采集乳品样品131份,阳性样品1份,检出率1.53%;肉制品224份,阳性样品4份,检测率1.79%。结论建立的单增李斯特氏菌检测PCR-免疫胶体金试纸条法特异度好,敏感度高,适用于食品中单增李斯特氏菌的检测。

ABSTRACT:Objective To establish the PCR-immunogold method for detection ofListeria monocytogenes in food.MethodsThe PCR method forListeria monocytogenes detection was built through the design of specific primers and the rapid test immunogold strip for detection of PCR products was established. Then the specificity and sensitivity of the new method were tested using the test bacterial strains andListeria monocytogenes in milk and meat products was detected using the PCR-immunogold strip method to verify whether this method is appropriate to food samples test.ResultsThe method established is specific and the sensitivity is higher than that of the agarose gel electrophoresis by 100 times. We screened 131 milk samples and 224 meat samples totally and found 1 (1.53%) of milk samples and 4 (1.79%) of meat samples carrying Listeria monocytogenes.ConclusionThe new method has high specificity and sensitivity and is suitable for the detection of Listeria monocytogenes in food.