人血清癌胚抗原的生物素-亲和素ELISA检测方法的建立

Establishment of BA-ELISA method for detecting CEA in human sera

目的:建立检测血清癌胚抗原(CEA)的高灵敏度生物素-亲和素酶联免疫检测(BA-ELISA)方法。方法:亲和层析纯化得到CEA,免疫新西兰家兔制备多克隆抗体。将得到的抗体连接生物素和辣根过氧化物酶,在生物素化BSA包被的96微孔板上建立生物素-亲和素ELISA(BA-ELISA)。对这一体系的线性、灵敏度、特异度、稳定性、回收率等参数进行鉴定,并和常规ELISA、放射免疫检测以及化学发光法相比较检测了临床标本中的CEA浓度。结果:线性检测范围为(0.42～50)U/mL,检测结果表明,体系稳定性良好;灵敏度为0.42 U/mL,批内、批间差异均小于10.0%。该研究建立的方法与常规ELISA对比差异有统计学意义,与放射免疫检测对比差别无统计学意义。与化学发光法相比较,回归方程为:y=0.04825+0.99674x,r=0.994,差异无统计学意义。结论:建立的CEA的BA-ELISA检测方法易于操作、灵敏度高、价格低廉,适合应用于临床检测。

AIM： To establish a sensitive biotin-avidin enzyme linked immunosorbent assay（BA-ELISA） method for detecting carcinoembryonic antigen（CEA） in serum.METHODS： CEA which had been purified by affinity chromatography was used to immunize the rabbits to produce polyclonal antibodies.Then the antibodies were connected with biotin and horseradish peroxidase（HRP）.So BA-ELISA method was established on the basis of 96 microwell plates coated with biotinylated BSA.Finally we examined the sensitivity,specificity,stability and recovery rate of this system and compared the BA-ELISA method with the traditional ELISA,radioimmunoassay and chemiluminescence in detecting CEA concentrations.RESULTS： The stability of this system was proved good.The linear range was from 0.42 to 50 U/mL,the sensitivity was 0.42 U/mL,and the intra-differences together with inter-differences were less than 10.0%.There was significant difference between BA-ELISA and traditional ELISA,while there was no significant difference between BA-ELISA and radioimmunoassay.The regression equation of this method was y=0.04825＋0.99674x and r=0.994,and there was no significant difference between the BA-ELISA and chemiluminescence.CONCLUSION： The BA-ELISA method we established to detect CEA was easy to operate,highly sensitive,low in price and suitable for application in clinical detection.