Kits for Auxiliary Diagnosis of Tuberculosis
A technology of auxiliary diagnosis and kit, applied in the direction of anti-animal/human immunoglobulin, instrument, anti-cytokine/lymphokine/interferon immunoglobulin, etc., can solve the problem of many affected factors
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Embodiment 1
[0027] Embodiment 1, the preparation of kit
[0028] The kit consists of antibody-coated microsphere mixture, biotin-labeled antibody mixture, PE-labeled streptavidin, and antigen stimulus.
[0029] 1. Antibody-coated microsphere mixture
[0030] Consists of 5 microspheres coated with different antibodies (the amount of each microsphere is equal); that is, microspheres coated with the following 5 antibodies: (1) IFN-γ antibody; (2) TNF-α antibody (3) IL-2 antibody; (4) MIG antibody; (5) IP-10 antibody. Each antibody-coated microsphere was purchased from Bender Medsystems, and the product numbers are listed in Table 1
[0031] Table 1 Product numbers of 5 kinds of microspheres coated with different antibodies
[0032] Microspheres coated with IFN-γ antibody
Human IFN-γ Simplex FlowCytomix, A4 beads
BMS8228FF
Microspheres coated with TNF-α antibody
Human TNF-α Simplex Flow Cytomix, B9 beads
BMS8223FF
Microspheres coated with IL-2 antibod...
Embodiment 2
[0046] Embodiment 2, the application of kit
[0047] Peripheral whole blood samples of 104 volunteers (58 cases of active tuberculosis patients and 46 cases of normal healthy people) were collected respectively, and tested with the kit prepared in Example 1. The detection method of each specimen is as follows:
[0048] (1) Separation of human peripheral blood mononuclear cells (PBMCs): Take 2ml of EDTA anticoagulated blood, and perform Ficoll density gradient centrifugation (room temperature 2000rpm, 20min) to obtain PBMCs. RPMI 1640 complete culture medium with 10% fetal bovine serum adjusted the cell density to 2×10 6 / mL, added to a 96-well cell culture plate (200 μl PBMC cell suspension per well).
[0049] (2) Antigen stimulation of PBMCs cells: Add 10 μl of antigen stimulator to each well, in 5% CO 2 , Cultivate in a 37°C incubator for 16-20 hours.
[0050] (3) Take 25 μl of the antigen-stimulated culture supernatant and add 25 μl of antibody-coated microsphere mixtur...
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