Tuberculosis infection T cell immune tetramer detection method

Abstract

The invention discloses a tuberculosis infection T cell immune tetramer detection method, peripheral blood mononuclear cells are separated from a whole blood sample, the peripheral blood mononuclear cells are incubated together with mycobacterium tuberculosis specific antigen, when effector T lymphocytes exist, TCR (T cell antigen receptor) in the mycobacterium tuberculosis specific antigen and MHC (major histocompability complex)-polypeptide compound are combined, with the aid of biotin-streptavidin system, MHC-polypeptide compound forms a tetramer molecule, single MHC-polypeptide tetramer molecule is combined with a plurality of TCR on the T cell surface to improve the MHC-polypeptide compound and TCR affinity and stability; a biotin labeled antibody mark detection PE (poly ethylene) fluorescent dye is added, a flow cytometer is used to count specific T cells for auxiliary diagnosis of tuberculosis, and the tuberculosis infection T cell immune tetramer detection method has the advantages of high in detection sensitivity, accuracy and product specificity, easy and simple application, safety and reliablility.

Description

technical field [0001] The invention relates to the field of life sciences, in particular to a tetramer detection method for T cell immunity against tuberculosis infection. Background technique [0002] At present, the commonly used detection methods for T cell immunity to tuberculosis infection include sputum smear examination and sputum culture. Detection methods, if these methods are used too frequently, there will be certain damage to the patient. Contents of the invention [0003] In order to solve the problem of too long period and poor safety of nuclear infection T cell immune detection, the present invention provides a detection method, the technical solution adopted is: isolate peripheral blood mononuclear cells, that is, PBMCs, from whole blood samples, and It is incubated with Mycobacterium tuberculosis-specific antigens. When there are effector T lymphocytes against Mycobacterium tuberculosis in peripheral blood, TCR in Mycobacterium tuberculosis-specific anti...

AI Technical Summary

Problems solved by technology

[0003] In order to solve the problem of too long period and poor safety of nuclear infection T cell immune detection, the present invention provides a detection method, the technical solution adopted is: isolate peripheral blood mononuclear cells, that is, PBMCs, from whole blood samples, and It is incubated with Mycobacterium tuberculosis-specific antigens. When there are effector T lymphocytes against Mycobacterium tuberculosis in peripheral blood, TCR in Mycobacterium tuberculosis-specific antigens binds to MHC-polypeptide complexes, with the help of biotin-streptavidin System, the MHC-peptide complex forms a tetramer molecule, and a single MHC-peptide tetramer molecule binds to multiple TCRs on the surface of T cells to improve the affinity and stability of the MHC-peptide complex and TCR; then add The biotin-labeled detection antibody is labeled with PE fluorescent dye; finally, the specific T cells are counted by flow cytometry for auxiliary diagnosis of tuberculosis