研究用特异性的出血性大肠杆菌O157:H7单克隆抗体包被处理的PCR管,对样品中的病原菌进行特异性免疫富集,以免疫捕获-PCR(Immunocaptured PCR,IC-PCR)管中捕获到的病原菌作为模板进行PCR扩增。结果显示免疫捕获-PCR对病原菌出血性大肠杆菌O157:H7检测灵敏度达到102CFU/mL,比直接PCR和ELISA提高了103倍;特异性验证实验证实,对其他10株食源性致病菌进行检测均无目标条带。该方法将免疫学和分子生物学检测结合起来,灵敏度高,特异性强,检测周期短,检测迅速,是适合检测一线进行快速检测的新方法。

PCR tubes were coated with the monoclonal antibody against hemorrhagic Escherichia coli O157 to capture the suspected bacteria in samples.PCR was carried out to detect the captured bacteria.Results showed that the sensitivity of this method was up to 102CFU / mL.Compared with direct PCR and the ELISA,the sensitivity of this method was increased by 1 000 times.The PCR test in the other 10 non-target strains showed negative results,which revealed that the immunocaptured PCR method had high specificity.This method combined immunology detection with molecular biology detection.The immunological enrichment and amplification of PCR happened in the same PCR tube.This method not only improved the detection sensitivity and accuracy,but also shortened the detection time.Also,it reduced the detection cost and pathogenic loss.This method is a new rapid detection method which is suitable for on-site test.