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目的:探究免疫磁分离免疫磁分离结合胶体金免疫层析法快速检测大肠杆菌O157∶H7的实验室效价。方法:将我院检验科保存的抗大肠杆菌O157∶H7单克隆抗体用胶体金标记为标记抗体,在利用抗大肠杆菌O157∶H7多克隆抗体制备胶体金免疫层析试纸条,用免疫磁分离分离待测样本后结合胶体金免疫层析法检测样本中的大肠杆菌O157∶H7,观察比较单纯的胶体金免疫层析法与免疫磁分离处理后再用胶体金免疫层析法检测大肠杆菌O157∶H7灵敏度和检出情况。结果:纯培养样品检测结果显示,单纯的胶体金免疫层析法检测结果在浓度为7.6×104 CFU/mL菌液呈弱阳性,表明检测最低浓度为7.6×104 CFU/mL。而免疫磁分离结合胶体金免疫层析法在浓度为7.6×103 CFU/mL既可检测为阳性。大肠杆菌O157∶H7培养基中37℃培养5h后,单纯的胶体金免疫层析法检测结果呈(-);免疫磁分离后再用试纸条检测,结果呈(±)。结论:免疫磁分离结合胶体金免疫层析法在检测大肠杆菌O157∶H7降低了细菌检出的底限浓度,提高了检出率。
OBJECTIVE: To explore the laboratory titer of E. coli O157:H7 by immunomagnetic separation and colloidal gold immunochromatography. Methods: The anti-colibacillin O157: H7 monoclonal antibody preserved in the laboratory of our hospital was labeled with colloidal gold as a labeled antibody, and colloidal gold immunochromatographic test strip was prepared by using anti-E.coli O157: H7 polyclonal antibody. Isolation and separation of samples to be tested with colloidal gold immunochromatography test samples of E. coli O157: H7 observed and compared pure colloidal gold immunochromatography and immunomagnetic separation and then colloidal gold immunochromatography detection of Escherichia coli O157: H7 sensitivity and detection of the situation. Results: The results of pure cultured samples showed that the results of simple colloidal gold immunochromatography assay showed weak positive at the concentration of 7.6 × 104 CFU / mL, indicating that the lowest detection concentration was 7.6 × 104 CFU / mL. The immunomagnetic separation combined with colloidal gold immunochromatography at a concentration of 7.6 × 103 CFU / mL can be detected as positive. E. coli O157:H7 medium was incubated at 37 ℃ for 5h, the results of pure colloidal gold immunochromatography showed (-); immunomagnetic separation and then test strips, the result was (±). Conclusion: Immunomagnetic separation combined with colloidal gold immunochromatography in detection of E. coli O157: H7 reduced the detection limit of bacterial concentration and increased the detection rate.