Autoantibody assay method for immunological mediated I type diabetes diagnosis

Abstract

The present invention relates to a self-body antibody detection method for immune mediated diabetes type A diagnosis, belonging to the field of biological medicine technology. Said invention includes the following steps: (1), using reverse transcriptive enzyme to make mRNA undergo the process of reverse transcription. Then using high fidelity DNA polymerase to respectively synthesize target DNA, using ligase and protein expression plasmid to make connection, making it be transformed into colibacillus to respectively express target protein; (2), using maltose-binding protein affinity chromatographic column and GST affinity chromotographic column to purify and separate out expression protein, dividing every purified expression protein into two portions; (3), coating antigen: making four antigens be mixed in the coating buffer solution, adding coating buffer solution in every enzyme-labeled well, standing still; (4), closing; and (5), using double-antigen sandwich method to determine antibody. Said invention can once detect any one or several kinds of antibodies of GAD65, IA2, Phogrin and ICA 69.

Description

Technical field [0001] The present invention relates to a detection method used in the field of biomedical technology, specifically, an autoantibody detection method used in immune-mediated diagnosis of type 1 diabetes. Background technique [0002] Diabetes type diagnosis is based on the cause of diabetes. Diabetes is divided into 4 types, namely type 1 diabetes, type 2 diabetes, gestational diabetes and other special types of diabetes. Their harm to patients and treatment methods are completely different. Diagnosis before treatment is very important for patients. Type 1 diabetes is also called youth-onset diabetes, because it often develops before the age of 35, accounting for less than 10% of diabetes. The insulin-producing cells of the pancreas in the patient's body have been damaged, and insulin is absolutely lacking. The patient needs to use insulin from the onset of the disease. Treatment and lifelong use. Because type 1 and type 2 diabetes not only have different causes, ...

AI Technical Summary

Problems solved by technology

Its disadvantage is that it can only target GAD in patients with type 1 diabetes 65 、IA 2 Although a single determination of multiple autoantibodies can predict and diagnose the disease to a certain extent, each antibody determination costs 100-150 yuan, and the diagnostic rate is relatively low. The cost of joint detection of two or more antibodies Expensive, with many operation steps and long time, it is not a small burden for the tester and the testee