【摘要】 目的制备抗人叶酸(FA)抗血清并应用异硫氰酸荧光素(FITC)系统开发新型非均衡竞争技术,建立可常规应用的定量检测血清FA的化学发光免疫分析(CLIA)法。方法 FITC-FA类似物、FA抗体-HRP依次加入包被有抗FITC抗体的化学发光板,形成FITC抗体-FITC-FA类似物-FA抗体-HRP的免疫反应复合物;并进行方法学评价,同时与非FITC检测系统及罗氏化学发光免疫分析系统检测结果进行比较。结果成功制备FA抗血清并建立基于FITC系统的非均衡竞争CLIA;经方法学评价,自研法的线性相关系数绝对值大于0.990 0,灵敏度1.21nmol/L,线性范围1.21~38.80nmol/L,批内变异系数小于5%,自研法定量检测性能优于非FITC系统;与罗氏检测系统结果有较好相关性(R=0.908 1)。结论建立的非均衡竞争定量检测血清FA的CLIA法,具有良好的检测灵敏度与特异性,可应用于常规检测。更多还原

【Abstract】 Objective To prepare anti-folic acid(FA)polyclonal antibody and develop a new non-balanced competing chemiluminescence analysis for clinical detection of FA.Methods Established the detection method by added FITC-FA-analogs and FAHRP-antibody in the light emitting plate,which coated with anti-FITC antibody,to form the immune response complex of FITC/antibody-FITC-FA-analogs/FA-antibody-HRP.Then methodology evaluation was performed to evaluate the method performance;and further compared the detecting results with non-FITC system detection system and Electrochemiluminescence system(Roche Elecsys 2010).Results The ELISA results showed that the prepared anti-FA antibodies can recognize serum FA specificly.The methodology evaluation indicated that the linear correlation coefficient of the standard curve was 0.990 0;the analytical sensitivity was1.21ng/mL;the range of linear detection was 1.21~38.80ng/mL;The coefficient variability of intra-assay was<5%,which was better than the results of non-FITC detection system;and the correlation coefficient was 0.908 1compared with the Elecsys-2010 detection system.Conclusion The established chemiluminescence immunoassay for human serum FA has a good sensitivity and specificity,and suitable for clinical serum FA quantitative detecting.更多还原