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目的:获得特异性检测复合前列腺特异性抗原(c-PSA)的单克隆抗体配对,建立基于化学发光c-PSA免疫定量试剂。方法:利用市购c-PSA抗原免疫6周龄雌性BALB/c小鼠,间接法ELISA差异筛选抗c-PSA、游离前列腺特异性抗原(f-PSA)、总前列腺特异型抗原(t-PSA)的抗体,抗体配对后化学发光定量检测血清中c-PSA蛋白。结果:获得抗体配对1A10/7D6-SAE,经c-PSA标准品及临床血清样本初步筛选,该抗体配对适用于基于化学法发光的免疫反应定量检测试剂的开发;血清阳性样本与阴性样本检测结果显示具有统计学意义(P<0.000 1);阳性样本定量结果与Siemens公司复合前列腺特异性抗原测定试剂盒(直接化学发光法)的相关系数为0.97;线性范围为0.1~100 ng/ml;检测灵敏度为0.005 ng/ml。结论:成功筛选获得特异性检测c-PSA的抗体配对,并开发c-PSA化学发光免疫定量试剂,其检测能力与国际主流试剂相当。
OBJECTIVE: To obtain monoclonal antibodies specific for the detection of complex prostate-specific antigen (c-PSA), and to establish a chemiluminescent c-PSA quantitative reagent. Methods: 6-week-old female BALB / c mice were immunized with commercially available c-PSA antigen. The indirect anti-c-PSA, f- PSA, t- PSA ) Antibody, the antibody paired chemiluminescence quantitative detection of serum c-PSA protein. Results: Antibody pairing 1A10 / 7D6-SAE was obtained and screened by c-PSA standard and clinical serum samples. The antibody pairing was suitable for the development of quantitative immunoassay reagents based on chemiluminescence. Serum positive samples and negative samples (P <0.0001). The correlation coefficient between the quantitative results of the positive samples and Siemens compound prostate specific antigen assay kit (direct chemiluminescence method) was 0.97; the linear range was 0.1-100 ng / ml; the detection Sensitivity is 0.005 ng / ml. CONCLUSION: The antibody screening for specific detection of c-PSA is successfully screened and the c-PSA chemiluminescence quantitative reagent is developed. Its detection ability is comparable to that of the international mainstream reagents.