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探讨胶体金免疫层析方法在感染铜绿假单胞菌肺癌患者快速床边检验的应用价值。方法:用分子生物学方法克隆OprF基因,原核系统诱导表达并纯化OprF,免疫BALB/c小鼠后通过杂交瘤技术制备特异性的mAb。采用双抗体夹心法,建立了检测铜绿假单胞菌的胶体金免疫层析法,并对该方法的特异性、灵敏度和稳定性进行了评价,对121例晚期肺癌患者临床标本进行了检测。结果:成功克隆OprF基因,经原核诱导表达获得铜绿假单胞菌的OprF。通过杂交瘤技术筛选,经鉴定单克隆抗体3C3B5可作为捕获抗体,多克隆抗体被HRP标记后可作为检测抗体。以传统细菌培养和鉴定方法为参照,用胶体金免疫层析试纸条检测的敏感度为83.3%(45/54),特异性为92.5%(62/67),总符合率为88.4%。结论:成功制备了铜绿假单胞菌胶体金免疫层析法试纸条,实现了晚期肺癌感染患者床边检验和病情检测。中华肿瘤防治杂志,2012,19(2):110-114
To investigate the application value of colloidal gold immunochromatography in the rapid bedside test of patients with Pseudomonas aeruginosa lung cancer. Methods: The OprF gene was cloned by molecular biology method. The prokaryotic expression system was purified and the OprF was purified. The specific mAb was prepared by the hybridoma technique after BALB / c mice were immunized. The double antibody sandwich method was used to establish the colloidal gold immunochromatography assay for detecting Pseudomonas aeruginosa. The specificity, sensitivity and stability of the method were evaluated. The clinical samples of 121 patients with advanced lung cancer were detected. Results: OprF gene was cloned successfully and OprF of Pseudomonas aeruginosa was obtained by prokaryotic expression. Through hybridoma technology screening, the monoclonal antibody 3C3B5 was identified as capture antibody, polyclonal antibody labeled with HRP can be used as detection antibody. The sensitivity and specificity of colloidal gold immunochromatographic test strip were 83.3% (45/54) and 92.5% (62/67), respectively. The total coincidence rate was 88.4%. Conclusion: The Pseudomonas aeruginosa colloidal gold immunochromatographic test strips were prepared successfully, and the bedside test and disease detection of patients with advanced lung cancer were achieved. Chinese Journal of Cancer Prevention and Treatment, 2012,19 (2): 110-114