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

Inactive Publication Date: 2011-12-28
程小星 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method is based on the secretion of a single cytokine, and many factors are affected. It has been reported that its sensitivity for diagnosing active tuberculosis is only 64

Method used

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  • Kits for Auxiliary Diagnosis of Tuberculosis
  • Kits for Auxiliary Diagnosis of Tuberculosis
  • Kits for Auxiliary Diagnosis of Tuberculosis

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a kit for assisted diagnosis of tuberculosis, which comprises a specific antibody composition, wherein the specific antibody composition comprises the following five antibodies: (1) IFN-gamma antibody, (2) TNF-alpha antibody, (3) IL-2 antibody, (4) MIG antibody and (5) IP-10 antibody. The kit disclosed by the invention can distinguish a Mycobacterium tuberculosis infected person from a BCG vaccinee. Compared with the ELISPOT tuberculosis diagnosis method, the tuberculosis diagnosis kit based on multi-molecular marker detection has obviously enhanced sensitivity to active tuberculosis (from 72% to 89%), and the positive rate for normal healthy persons is obviously reduced (from 27% to 16%).

Description

technical field [0001] The invention relates to a kit for auxiliary diagnosis of tuberculosis. Background technique [0002] There are still many problems in the diagnosis of tuberculosis. Sputum smear acid-fast staining has relatively high specificity in the diagnosis of pulmonary tuberculosis, but the sensitivity is low, about 15%-30%. The cultivation of Mycobacterium tuberculosis takes 2-6 weeks, and the technique is complex, expensive and time-consuming. The PCR-based method has the characteristics of simplicity, rapidity and high sensitivity, but the result is affected by many factors and has poor stability, so it should not be used alone for the diagnosis of tuberculosis. X-ray examination has important reference value in the diagnosis of tuberculosis, but in many cases it is difficult to differentiate it from other diseases, especially atypical lesions. [0003] Serological diagnostic methods (such as ELISA) have the advantages of being simple and fast, and are eas...

Claims

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Application Information

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IPC IPC(8): G01N33/569C07K7/08C07K14/35C07K16/24
Inventor 程小星王心静蒋静曹志红
Owner 程小星
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