Establishment Of Chemiluminescence Method And Its Application In The Diagnosis Of Tuberculosis

TB (tuberculosis, TB) is an infectious disease caused by Mycobacterium tuberculosis(mycobacterium-tuberculosis, MTB) caused clinical diagnosis of tuberculosis is mainlydivided into three categories: bacteriology, molecular biology and immunology check, whichimmunological tests with high accuracy, easy operation and concern. In recent years,immunological tests in γ-interferon (IFN-γ) release assay (Interferon Gamma Release Assay,IGRA) due to the high sensitivity and specificity aroused interest. ELISPOT-based IGRA test(ELISPOT) the first to be widely used, the World Health Organization (WHO) recommendstwo QFT-G TB diagnostic reagents kits and T-SPOT.TB kits are based ELISPOT.IGRA is the use of enzyme-linked immunosorbent assay (ELISA) method forquantitative determination of IFN-γ release in whole blood in the stimulation of M.tuberculosis specific antigens levels for the diagnosis of tuberculosis and Mycobacteriumtuberculosis infection. Detection principle is to use the principle of sandwich method fordetecting, measuring antigen-specific cell-mediated immune response intensity. Stimulationby Mycobacterium tuberculosis infection by M. tuberculosis-specific antigens specific to theproliferation of T lymphocytes and release IFN-γ. With IFN-γ antibody coated magneticparticles conjugate, horseradish peroxidase-labeled IFN-γ antibody enzyme. Formation ofantibodies immunoreactive-antigen-antibody complexes, the catalytic composite photonicissued luminescent substrate, and the luminous intensity is proportional to the content ofIFN-γ.In this study, based on the establishment of a chemiluminescent IGRA method, andapply it to the diagnosis of tuberculosis. The suspension was prepared by chemiluminescencemethod magnetic particles, conjugate preparation, preparation of calibration products,optimize the reaction system, the normal reference values determined from stability studies toestablish. The method is applied to the diagnosis of tuberculosis, mainly through researchand clinical diagnosis information comparing chemiluminescence method is applied to theassessment of TB diagnostic sensitivity, specificity and accuracy of indicators to OxfordImmunotec Ltd production of T-SPOT.TB kit as a control, chemiluminescence method wasapplied to the assessment and diagnosis of TB-positive T-SPOT.TB kit compliance rate, negative coincidence rate and total compliance rate. Chemiluminescence detectionestablished shortened to30minutes, Clinical diagnosis information and Oxford ImmunotecLtd produces T-SPOT.TB kit as a control, analysis of test results with clinical diagnosis,assessment of the performance of the reagents results were compared with the clinicaldiagnosis, the sensitivity was86.19%(69/81) and a specificity of84.26%(589/699), theaccuracy was84.36%(658/780). And T-SPOT.TB kit for benchmarking, the positive rate was85.88%(225/262), the rate was81.44%(215/264), total coincidence rate83.65%(440/526).This study established chemiluminescence method used in the diagnosis of tuberculosis,shortening the detection time and improve the linear range, more sensitive and specific, andno environmental pollution. TB diagnosis using chemiluminescence method to lay thefoundation for future.