Evaluation and application of double antibody sandwich ELISA kit for micro-quantitative soluble complement receptor 1 protein in human serum / 国际检验医学杂志

ABSTRACT

Objective To evaluate the repeatability and stability of double antibody sandwich ELISA kit for micro-quantitative soluble complement receptor 1(sCR1) in human serum and to understand its practical application effect .Methods 50 patients with middle and advanced liver cirrhosis and 50 individuals undergoing physical examination served as the liver disease group and normal control group respectively .The mouse anti-human CD35 monoclone antibody ,rabbit anti-human sCR1 polyclonal antibody and goat anti-rabbit IgG labeled by horseradish peroxidase served as the envelope antibody ,sandwich antibody and detection antiboby .The purified recombinant human sCR1 protein served as the standard substance .The human serum micro-quantitative double antibody sandwich CR1 ELISA kit was established .Then the repeatability and stability tests were performed .Then the sCR1 protein level of two group of serum was detected by this kit .Results The linear range of double antibody sandwich ELISA for detecting human se-rum micro-quantitative sCR1 protein was 15 .60 -250 .00 ng/mL ;the regression equation of sCR1 protein concentration to absor-bance value was Y=112 .10X2 +18 .21X+1 .694(r2 =0 .998);in the repeatability test ,the intra-batch relative standard deviation (RSD) in high and low concentrations of standard substance detection value was 6 .20% and 7 .40% respectively ,the inter-batch RSD was 6 .70% and 7 .90% respectively ;in the stability test ,RSD was not more than 0 .01;the serum sCR1 expression level in the liver disease group was significantly higher than that in the normal control group (P<0 .01) .Conclusion The human serum double antibody sandwich ELISA kit for detecting human sCR1 has wide linear range ,good repeatability ,is easy to be stored and suitable for clini-cal and scientific research detection work .