A novel electrochemiluminescence method for detecting insulin autoantibody

Zhonghua yi xue za zhi
Z WangZ G Zhou

Abstract

Objective: To establish the electrochemiluminescence (ECL) method for insulin autoantibody (IAA) detection in China and preliminarily evaluate its application value. Methods: Proinsulin was labeled with Sulfo-tag and biotin, and then incubated with IAA in serum. The Meso Scale Discovery (MSD) streptavidin plate was used to capture the labeled antigen-antibody complex, which was subsequently detected by MSD electrochemiluminescence machine. IAA index ≥ 0.005 was used as a positive threshold. After optimizing the detection conditions, the correlation and consistency of ECL method and microplate radioimmunoassay were compared. Patients with type 1 diabetes mellitus (T1DM, n=55) and first-degree relatives (n=216) from the Department of Metabolism and Endocrinology of the Second Xiangya Hospital of Central South University from 2016 to 2018, and healthy volunteers (n=413) were selected as subjects, and their serum samples were tested to evaluate the clinical application value of ECL. Results: The optimized conditions included as follows: (1) The optimal concentration of biotin-proinsulin and Sulfo-tag-proinsulin was 800 ng/ml. (2) Serum acidification was achieved by incubating for 45 min. (3) Adding 35 μl diluted antigen, followed b...Continue Reading

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