摘要
目的:建立检测甲胎蛋白化学发光免疫定量分析方法.方法:采用双抗体异位点一步夹心法,以甲胎蛋白单克隆抗体作为固相包被,采用改良过碘酸钠法进行甲胎蛋白多克隆抗体与碱性磷酸酶偶联制备酶标抗体;以金刚烷胺衍生物CSPD作为底物,优化CSPD与SapphireⅡ发光体系;用国家标准品校定定量标准品,建立了人血清AFP的化学发光免疫定量分析法.用该法检测800份正常人血清,确定正常值参考范围,将临床血清标本100份与bayer Acs:180(全自动发光体系统进行比较.结果:本方法的最低检出量为2.0ng/mL,线性范围2.0～600.0ng/mL,批内和批间的变异率分别为6.7%和7.7%,回收率在98.2%～102.5%之间.与CEA(3μg/mL)的交叉≤0.15%;与铁蛋白(10μg/mL)≤0.04%;与人血清白蛋白(200g/L)≤2.5×10-8,与人血清丙球蛋白(100g/L)≤2.0×10-8,与bayerAcs:180(比较相关系数r=0.994(P<0.001).结论:建立的甲胎蛋白化学发光免疫分析法可常规用于临床检测.
AIM： To establish a method of chemiluminescent immunoassay for determination of alpha -fetoprotein （ AFP ） in human serum. METHODS： The two-site enzyme immunoassay is based on the direct sandwich technique. A monoclonal antibody was bonded to the microplate for coating and the conjugation of alkaline phosphatase to another polyclonal antibody was performed by NaIO4 method. Chemiluminescent immunoassay was established by optimizing CSPD and SapphireⅡ luminescent system. Sensitivity, linear scope, precision, interference and recovery for the method were evaluated. The reference range was defined by testing 800 serum samples from healthy subjects. The comparison for 100 serum samples from patients was assayed by the bayer Acs： 180 Automated Immunoassay system. RESULTS ： The detectable minimum was 2.0 ng/mL. The linear scope was from 2.0 to 600.0 ng/mL. Average inter and intra-assay were 7.7% and 6.7% , respectively. The recoveries ranged from 98.2% to 102.5%. The cross-reacting rate for CEA（3 μg/mL）, Ferritin （10 μg/ml）, HAS（200 g/L）, IgG（100 g/L） were≤〈0.15%, 0. 04%, 2.5 × 10^-8 and 2.0 × 10^-8, respectively. Compared with bayer Acs：180 Automated immunoassay system, the relative coefficient was 0. 994 （ P 〈 0. 001 ）. CONCLUSION： A successful establishment of chemiluminescent immunoassay provides a way for clinical determination of AFP.
出处
《第四军医大学学报》
CAS
北大核心
2007年第14期1291-1293,共3页
Journal of the Fourth Military Medical University
关键词
甲胎蛋白类
化学发光测定法
定量分析
alpha-fetoproteins
chemiluminescent measurements
quantitative analysis
