Nonradioactive detecting methods of inflammatory myopathy HMGCR (3-hydroxy3-methylutaryl coenzyme A reductase) autoantibody and application

Abstract

The invention discloses nonradioactive detecting methods of an inflammatory myopathy HMGCR (3-hydroxy3-methylutaryl coenzyme A reductase) autoantibody and an application. The nonradioactive detecting methods comprise a nonradioactive labeled immunoprecipitation assay method and an immunoblotting assay method of the inflammatory myopathy HMGCR autoantibody and is used for detecting the HMGCR-autoantibody of a patient with IIM (idiopathic inflammatory myopathies), wherein the nonradioactive labeled immunoprecipitation assay method of the inflammatory myopathy HMGCR autoantibody comprises the steps as follows: Protein G magnetic beads are conjugated with 20 mu l of serum of an HMGCR antibody-positive patient with inflammatory myopathy or 20 mu l of healthy control serum respectively, the serum is then incubated with a cell lysate overexpressing HMGCR C terminal for antigen adsorption, SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) is performed after elution, and an c-myc antibody is used for detection; the immunoblot assay method of the inflammatory myopathy HMGCR autoantibody comprises the step that the HEK 293 cell lysate overexpressing the HMGCR C terminal is subjected to SDS-PAGE and detected by the serum of the HMGCR antibody-positive patient as a primary antibody and a rabbit-anti-human IgG H & L as secondary antibody. With the adoption of the nonradioactive detecting methods, the problems of the lack of commercial ELISA kits and immunoblotting membrane strips for detecting the HMGCR antibody at home, as well as high cost and false positive rate and low safety of radiolabeled immunoprecipitation of an ELISA kit self-coated with an HMGCR antigen are solved.

Description

technical field [0001] The application belongs to the field of biotechnology, and relates to a non-radioactive detection method and application of HMGCR autoantibodies in inflammatory myopathy, in particular to a non-radioactive labeling immunoprecipitation detection method and immunoblotting detection method and application of HMGCR autoantibodies in inflammatory myopathy. Background technique [0002] Idiopathic inflammatory myopathy (idiopathic inflammatory myopathies, IIM) is a group of systemic autoimmune diseases mainly invading skeletal muscle, the abnormality of autoimmunity is the key to the occurrence and development of IIM. According to the latest diagnostic classification criteria, IIM can be divided into polymyositis (PM), dermatomyositis (DM), immune-mediated necrotizing myopathy (IMNM), nonspecific Myositis (nonspecific myositis, NSM) and inclusion body myositis (inclusion body myositis, sIBM). The clinical features of IIM are muscle weakness in the proximal ...

AI Technical Summary

Problems solved by technology

[0007] In order to overcome the shortcomings of the above-mentioned prior art, solve the high cost, high false positive rate and radioactivity of ELISA kits and western blot membrane strips for the detection of HMGCR antibodies that are not yet commercialized in China, as well as self-coated ELISA kits for HMGCR antigens. To address the safety issues of the immunoprecipitation method, the purpose of the present invention is to provide a non-radioactive detection method for HMGCR autoantibodies in inflammatory myopathy, specifically including a non-radioactive detection method for HMGCR autoantibodies in inflammatory myopathy and a Western blot detection method

Images