PCR-免疫胶体金试纸条方法检测食品中肠出血性大肠杆菌O157:H7

摘要

目的：建立PCR-免疫胶体金试纸条法快速检测食品中肠出血性大肠杆菌O157：H7的分析方法。方法通过设计特异性引物建立肠出血性大肠杆菌O157：H7 PCR检测方法并使用免疫胶体金技术以及双抗体夹心法建立PCR产物快速检测试纸条并设计核酸检测展开液；将1株大肠杆菌O157： H7标准菌株和7株其他常见食源性致病菌作为试验菌株，用试验菌株检测PCR-免疫胶体金试纸条方法的检测特异度，并比较PCR-免疫胶体金试纸条法和PCR-琼脂糖凝胶电泳法的检测敏感度。结果 PCR-免疫胶体金法具有良好的特异度，灵敏度比标准琼脂糖凝胶电泳法高100倍。结论本文建立的肠出血性大肠杆菌 O157：H7检测 PCR-免疫胶体金试纸条法特异度好，灵敏度高，价格低廉，适用于食品中肠出血性大肠杆菌O157：H7的检测。%ABSTRACT:Objective To establish a PCR-immunogold strip method for rapid detection ofEscherichia coli O157:H7 in food.MethodsThe PCR method was established through the design of specific primers and the establishment of rapid test immunogold strip for detection of PCR products combined immunogold skills with double antibody sandwich method. Test specificity of the new method using the test bacterial strains which include 1Escherichia coli O157:H7 and 7 common food-borne pathogens and compare the sensitivities of the immunogold strip method and the traditional agarose gel electrophoresis method.ResultsThe established method was specific and the sensitivity was higher than the agarose gel electrophoresis by 100 times. ConclusionThe new method is suitable for the detection ofEscherichia coli O157:H7 in food.