Method for detecting human myocardial troponin T through cytometric bead array

Abstract

The invention discloses a method for detecting human myocardial troponin T through cytometric bead array. The method comprises the following three steps of: activating a bead, coupling a monoclonal antibody and detecting the human myocardial troponin T, specifically, firstly, activating the beads; secondly, preparing the monoclonal antibody of the carboxylation bead coupled mouse anti human myocardial troponin T; thirdly, capturing the human myocardial troponin T in a specimen; and finally, adding multiple antigenic and fluorescein isothiocyanate labeled donkey anti goat intravenous gamma globulin G of goat anti human myocardial troponin T, and detecting the fluorescence intensity of fluorescein isothiocyanate (FITC) on the bead on a flow cytometry to indirectly detect the content of the human myocardial troponin T in the specimen. The monoclonal antibody coupled bead, the polyclonal antibody of the goat anti human myocardial troponin T, and the fluorescein isothiocyanate labeled donkey anti goat intravenous gamma globulin (IgG) can be prepared into a commodity kit, which can detect the human myocardial troponin T at any time. The method is simple and feasible; and by the method, the human myocardial troponin T can be quickly detected with convenience and the basis for disease diagnosis is provided.

Description

technical field [0001] The invention belongs to a method for analyzing biological substances, in particular, it belongs to a flow microsphere analysis technology method in flow cytometry analysis. Background technique [0002] Cardiac troponin is currently the biomarker with the highest specificity and high sensitivity in the diagnosis of myocardial injury and necrosis, and it also has important application value in the risk stratification of acute coronary syndrome (ACS). The "gold standard" of necrosis. At present, the methods for detecting human cardiac troponin T mainly include: 1. Immunogold standard method, which can be quickly qualitative or semi-quantitative; 2. Electrochemiluminescence method, which is sensitive (minimum detection limit 10pg / mL) and precise, but reagent costs 3. Immunoturbidimetry is practical, convenient, fast, and low in cost, but there are many influencing factors, such as excessive antigen or antibody, soluble complexes may appear, causing erro...

AI Technical Summary

Problems solved by technology

At present, the methods for detecting human cardiac troponin T mainly include: 1. Immunogold standard method, which can be quickly qualitative or semi-quantitative; 2. Electrochemiluminescence method, which is sensitive (minimum detection limit 10pg / mL) and precise, but reagent costs 3. Immunoturbidimetry is practical, convenient, fast, and low in cost, but there are many influencing factors, such as excessive antigen or antibody, soluble complexes may appear, causing errors, and The blood lipid content affects the turbidity, resulting in a false increase, which is not conducive to the unification and comparison of test methods; other methods, such as radioimmunoassay and enzyme immunoassay, have poor practicability

At present, there are many clinical methods for detecting human cardiac troponin T, and there are also some patented technical solutions, such as "Human myoglobin / creatine kinase isoenzyme / cardiac troponin I diagnostic test paper" No. I color particle diagnostic test paper", No. 200780010683.7 "Tools and methods for distinguishing acute and chronic myocardial necrosis in symptomatic patients", No. 200810036418.2 "Troponin I serum rapid detection kit (colloidal gold method)", etc., but all There are standardization issues