Kit for detecting active tuberculosis based on antigen-specific TNF-alpha-ELISA (enzyme linked immunosorbent assay) and application thereof

A kit and specific technology, applied in biological testing, measuring devices, disease diagnosis, etc., can solve the problems of similar products that have not been developed for the diagnosis of active tuberculosis

Inactive Publication Date: 2014-02-26
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no similar product developed for the diagnosis of active tuberculosis in China

Method used

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  • Kit for detecting active tuberculosis based on antigen-specific TNF-alpha-ELISA (enzyme linked immunosorbent assay) and application thereof
  • Kit for detecting active tuberculosis based on antigen-specific TNF-alpha-ELISA (enzyme linked immunosorbent assay) and application thereof
  • Kit for detecting active tuberculosis based on antigen-specific TNF-alpha-ELISA (enzyme linked immunosorbent assay) and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] A test kit for detecting active tuberculosis based on TNF-α-ELISA, the test kit comprising:

[0047] 1) ELISA plate coated with TNF-α antibody;

[0048] 2) TNF-α protein standard;

[0049] 3) Protein solution of ESAT-6 and CFP-10;

[0050] 4) Biotin-labeled TNF-α antibody;

[0051] The kit also includes any one or more of PBS concentrated washing solution, phytohemagglutinin, substrate solution and stop solution.

[0052] The kit also includes avidin-labeled horseradish peroxidase.

[0053] The protein solution concentrations of ESAT-6 and CFP-10 were both 10 μg / ml.

[0054] .The application of the kit in detecting active tuberculosis comprises the following steps:

[0055] 1) Add 100 μl of peripheral blood mononuclear cell suspension to be tested into four wells of a common 96-well reaction plate;

[0056] 2) Add the following reagents to the above four wells: add 50 μl culture solution to the negative control well; add 50 μl phytohemagglutinin to the positive co...

Embodiment 2

[0072] 20 cases of active tuberculosis patients, 20 cases of latent tuberculosis controls, 10 cases of non-tuberculosis patients and 10 cases of healthy controls were tested using the above kits and corresponding methods.

[0073] A case selection

[0074] All patients and controls were from Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology. The inclusion criteria for the four groups of people are as follows:

[0075] Active tuberculosis patients: clinical symptoms such as fever, night sweats or weight loss, and positive clinical smear or culture.

[0076] Latent tuberculosis control subjects: no clinical symptoms such as fever, night sweats or weight loss, no imaging manifestations of tuberculosis, but positive tuberculosis infection T cell detection (T-SPOT).

[0077] Non-tuberculosis patients: have clinical symptoms such as fever, night sweats or weight loss, but the T-cell test for tuberculosis infection (T-SPOT) is negat...

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Abstract

The invention discloses a kit for detecting active tuberculosis based on antigen-specific TNF-alpha-ELISA (enzyme linked immunosorbent assay) and an application thereof. The kit comprises an ELISA plate coating a TNF-alpha antibody, a TNF-alpha protein standard, protein liquid of ESAT-6 and CFP-10, a biotin-marked TNF-alpha antibody and avidin-marked horseradish peroxidase. By adopting the kit for detecting active tuberculosis based on TNF-alpha-ELISA, a method for distinguishing active tuberculosis infection from latent tuberculosis infection is established; the TNF-alpha released by the peripheral blood mononuclear cell under the stimulus of tuberculosis-specific antigens ESAT-6 and CFP-10 and the background TNF-alpha released by the peripheral blood mononuclear cell without stimulus are quantitatively detected through the ELISA technology, and the value of the antigen-specific TNF-alpha is obtained by subtracting the two so as to perform direct diagnosis on the active tuberculosis.

Description

technical field [0001] The invention relates to the application field of ELISA kits, in particular to a kit for detecting active tuberculosis based on antigen-specific TNF-α-ELISA and its application. Background technique [0002] Tuberculosis is one of the major diseases that threaten human health. It mainly causes lung diseases, and its morbidity and mortality are high. In history, tuberculosis was widely prevalent in the world, killing hundreds of millions of people. Since the 1950s, anti-tuberculosis drugs have been discovered continuously, and the prevalence of tuberculosis has been controlled to a certain extent. In recent years, due to the neglect of tuberculosis prevention and control work in some countries, coupled with factors such as the increase of floating population, the spread of HIV-infected people and the continuous emergence of drug-resistant tuberculosis strains, the prevalence of tuberculosis has rebounded. At present, there are 8 million new cases ever...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68G01N2800/12
Inventor 孙自镛汪峰侯红艳
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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