A gamma delta T cell surface activation molecule and kit for rapid diagnosis of active tuberculosis

A rapid diagnosis and kit technology, applied in analytical materials, biological tests, material testing products, etc., can solve problems such as low positive rate, high missed diagnosis rate, and inability to meet clinical needs, and achieve high sensitivity and specificity.

Active Publication Date: 2017-04-12
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, any single Mtb antigen cannot detect all antibodies in TB patients
Using this method alone has the defects of low positive rate and high missed diagnosis rate, which cannot meet the clinical needs

Method used

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  • A gamma delta T cell surface activation molecule and kit for rapid diagnosis of active tuberculosis
  • A gamma delta T cell surface activation molecule and kit for rapid diagnosis of active tuberculosis
  • A gamma delta T cell surface activation molecule and kit for rapid diagnosis of active tuberculosis

Examples

Experimental program
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Effect test

Embodiment 1

[0018] Example 1 Establishment of T cell surface molecule kit for rapid diagnosis of active tuberculosis

[0019] The kit of T cell surface molecules for rapid diagnosis of active tuberculosis includes the following components: (1) specific fluorescent antibody for TREM-1 (red); (2) specific fluorescent antibody for Vδ2TCR (green); (3) Fluorescent antibody for isotype control; (4) Reaction buffer.

[0020] Wherein, the reaction buffer is 1×PBS with 1% bovine serum albumin (BSA);

[0021] The concentrations of TREM-1 and Vδ2 TCR-specific antibodies are both 0.5mg / ml;

[0022] The isotype control antibody is the same type and the same fluorescently labeled IgG as the TREM-1 antibody;

[0023] (Negative) isotype control is the addition of Vδ2TCR antibody and isotype control antibody.

[0024] Specifically include the following steps:

[0025] (1) Use lymphocyte separation medium to separate PBMC, take 1×10 5 ~1×10 7 1 cell, add 1ml of reaction buffer, and centrifuge at 1500...

Embodiment 2

[0031] The detection rate of embodiment 2 clinical small sample verification kit

[0032] Peripheral blood was collected from subjects, including 42 healthy subjects and 76 active tuberculosis patients. Peripheral blood mononuclear cells (PBMCs) were isolated. Adopt the kit of the present invention to detect.

[0033] The kit of T cell surface molecules for rapid diagnosis of active tuberculosis includes the following components: (1) specific fluorescent antibody for TREM-1; (2) specific fluorescent antibody for Vδ2TCR; (3) fluorescent antibody for isotype control; (4) Reaction buffer.

[0034] Wherein, the reaction buffer is 1×PBS with 1% bovine serum albumin (BSA);

[0035] The concentrations of TREM-1 and Vδ2 TCR-specific antibodies are both 0.5mg / ml;

[0036] The isotype control antibody is the same type and the same fluorescently labeled IgG as the TREM-1 antibody;

[0037] (Negative) isotype control is the addition of Vδ2TCR antibody and isotype control antibody.

...

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Abstract

The invention discloses a gamma delta T cell surface-active molecule and a kit for quickly diagnosing active tuberculosis. According to the gamma delta T cell surface-active molecule and the kit disclosed by the invention, a specific T cell surface molecule is applied to the diagnosis of active tuberculosis for the first time; the gamma delta T cell surface-active molecule comprises a myeloid cell induced receptor 1 and / or a V delta 2T cell receptor; by using the two cell surface molecules and corresponding kits thereof, quick detection of active tuberculosis can be realized; the detection method and the detection kit of active tuberculosis have the advantages of high sensitivity, good specificity, simple operation, simplicity, convenience, quickness and the like; meanwhile, the detection method and the detection kit are low in price and are suitable for popularization and application.

Description

technical field [0001] The invention specifically relates to a gamma delta T cell surface activation molecule and a kit for rapidly diagnosing active tuberculosis. Background technique [0002] The gold standard for TB diagnosis is bacteriological examination, including acid-fast staining of sputum smear and isolation and culture of Mycobacterium tuberculosis. However, among tuberculosis patients, the detection rate of Mycobacterium tuberculosis is only 30%, which means that most of the patients are bacillus-negative tuberculosis, and it is difficult to obtain a diagnosis from etiology. At present, the laboratory diagnosis of bacillary negative tuberculosis largely relies on immunological detection methods. Traditional tuberculosis immunological detection methods include tuberculin test (TST) and serum tuberculosis antibody detection. TST uses tuberculin pure protein derivative (PPD) as an antigen to detect whether the body has been infected with Mycobacterium tuberculosis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/50
CPCG01N33/505G01N33/5695
Inventor 黄曦吴永坚
Owner SUN YAT SEN UNIV
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