血管紧张素Ⅰ化学发光免疫分析方法的建立

Establishment of chemiluminescence immunoassay for AngiotensinⅠ

目的：建立人血管紧张素Ⅰ（AngⅠ）化学发光免疫分析方法,通过测定血浆中血管紧张素Ⅰ的含量计算肾素活性。方法：用生物素标记AngⅠ抗原,荧光素标记AngⅠ抗体,将抗荧光素抗体包被于化学发光板,以链亲和素酶为催化剂,采用竞争法建立AngⅠ化学发光免疫分析方法。结果：本方法的测量范围为100～7 800 pg/ml,灵敏度为24.3 mU/ml,批内变异为5.6%～8.7%,批间变异为6.8%～10.4%,回收率为95.4%～105.3%,稀释实验测定值与理论值呈线性相关,相关系数为r=0.999。与放射免疫分析试剂盒的相关性方程分别为y=0.95x＋235.70,相关系数r=0.973。包被板、生物素标记AngⅠ抗原以及荧光素标记AngⅠ抗体在37℃存放一周稳定性良好。结论：方法学鉴定结果符合免疫分析的基本要求。

Objective：The chemiluminescence immunoassay for AngiotensinⅠ （ AngⅠ ） was developed by the competition method. The renin activity was calculated by the determination of Ang Ⅰ. Methods： AngⅠ antigen was labeled with biotin and fluorescein labeled AngⅠantibody,and anti fluorescein antibody was used to coated with the microporous plate . Results：The standard range of the method was 100-7 800 pg/ml. The assay sensitivity was 24. 3 pg/ml. The intra- and inter-assay coefficients of variance were 5. 6%-8. 7% and 6. 8%-10. 4% respectively. Analytical recovery was 95. 4%-105. 3%. The correlation coefficients between measured and expected values were 0. 999 after serial diluted. Compared with radioimmunoassay kit,the correlative equation was y=0. 95x＋235. 70,correlation coefficient was 0. 973. Coated microporous plate and biotin labeled Ang Ⅰ antigen,and fluorescein labeled Ang Ⅰantibody at 37℃ for a week with good stability. Conclusion： The results of the method were in accord with the basic requirements of immunoassay.