建立非均衡竞争叶酸定量测定的化学发光免疫分析法

发布时间：2018-05-08 17:28

  本文选题：FITC + 非均衡竞争　； 参考：《重庆医学》2017年06期

【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。
[Abstract]:Objective to prepare anti-folate FAA antiserum and develop a novel disequilibrium competitive technique by using fluorescein isothiocyanate (FITC) system, and to establish a CLIA-chemiluminescent immunoassay for quantitative detection of serum FA. Methods FITC-FA analogue FA antibody -HRP was added to chemiluminescence plate coated with anti FITC antibody in turn to form an immunoreactive complex of FITC antibody FITC-FA analogue -FA antibody -HRP, and the methodology was evaluated. The results were compared with non-FITC detection system and Roche chemiluminescence immunoassay system. Results the FA antiserum was successfully prepared and the unbalanced competitive CLIA based on FITC system was established. The linear correlation coefficient was more than 0.990, the sensitivity was 1.21 nmol / L, the linear range was 1.21 ~ 38.80 nmol / L, the coefficient of variation was less than 5, the self-developed quantitative detection performance was better than that of the non-FITC system, and there was a good correlation with the results of Roche detection system. Conclusion the CLIA method for the quantitative detection of serum FA has good sensitivity and specificity and can be used for routine detection.
【作者单位】： 重庆医科大学附属第二医院检验科;博奥赛斯(天津)生物科技有限公司;
【分类号】：R446.6
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本文编号：1862327