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Gold-labeled diagnosis reagent based on combined protein for tubercle bacillus

A technology of Mycobacterium tuberculosis and combined protein, which is applied in the field of medical immunodiagnosis, can solve the problems such as the need to improve the sensitivity and specificity, the long time and the poor stability, so as to improve the sensitivity and specificity, sensitivity and specificity. Improve and facilitate the effect of clinical promotion and application

Inactive Publication Date: 2008-07-09
THE 309TH HOSPITAL OF CHINESE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Difficult to distinguish tuberculosis from environmental mycobacterial infection and BCG vaccination positive conversion
2. The ELISA method needs to be operated by a special person. Different people operate with different batches of reagents, the results are very different, the stability is poor, and the time is also long, it takes 2-3 hours
The commercially available tuberculosis serum detection reagent IC-TB card contains PstS-1 intact protein, and its sensitivity and specificity need to be improved; other proteins have not been reported as serological detection reagents

Method used

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  • Gold-labeled diagnosis reagent based on combined protein for tubercle bacillus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Detection of Mycobacterium tuberculosis antibody by gold standard diafiltration antibody detection reagent

[0022] Take a gold standard diafiltration plate, add 2-4 drops of blocking solution, after the blocking solution is completely dry, add 2-4 drops of patient serum, after the serum is completely dry, add 2-4 drops of washing buffer, wait for washing After the buffer is completely dry, add 2-4 drops of gold-labeled secondary antibody. After the secondary antibody is dry, add 2-4 drops of washing buffer and wait for 5-10 minutes. After the left quality control point is completely colored, observe the right Whether the dot is colored, if it is red, it is a positive result, and if there is no dot, it is a negative result.

Embodiment 2

[0023] Embodiment 2: Gold standard diafiltration (silver enhanced) antibody detection reagent detects Mycobacterium tuberculosis antibody

[0024] Take a gold standard diafiltration plate, add 2-4 drops of blocking solution, after the blocking solution is completely dry, add 2-4 drops of patient serum, after the serum is completely dry, add 2-4 drops of washing buffer, wait for washing After the buffer is completely dry, add 2-4 drops of gold-labeled secondary antibody. After the secondary antibody is dry, add 2-4 drops of washing buffer and wait for 5-10 minutes. After the quality control point on the left is completely colored, add silver Add 2-4 drops of the strengthening reagent, and after 5-10 minutes, observe whether the dot on the right is colored. If it is black, it is a positive result, and if there is no dot, it is a negative result.

[0025] Effect experiment of the present invention is as table 1

[0026] Table 1 Different reagents detect the positive rate of diff...

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Abstract

The invention discloses a medical immunity diagnose technique, especially providing a bacillus tubercle mycobacterium gold mark diagnose agent based on combined protein. Wherein, it uses B cell antigen determinants of bacillus tubercle mycobacterium main excrete proteins as ESAT6, MPT64, PstS-1, Ag85B to combine a new group of protein as antigen, as the specific test antibody in the clinic serology diagnosis of phthisis. The invention combines the B cell antigen determinants of bacillus tubercle mycobacterium main excrete proteins to be displayed by gene project, and purified to attain one new protein, and via gold mark filter method to test the serum of tuberculosis patient. The invention has high sensitivity, simple operation, and high speed, which can identify the sensitized condition caused by contacting the non- bacillus tubercle mycobacterium or inoculating beg vaccine, and the real bacillus tubercle mycobacterium. It can be used in the clinic serology diagnosis of clinic tuberculosis.

Description

technical field [0001] The invention belongs to the technical field of medical immunodiagnosis, and relates to a reagent for diagnosis and detection of Mycobacterium tuberculosis using a new protein composed of ESAT6, MPT64, PstS-1 and Ag85B antigenic determinants mainly secreted by Mycobacterium tuberculosis and its application in clinical tuberculosis application in diagnosis. Background of the invention [0002] Currently, tuberculosis is on the rise globally due to the rise of drug-resistant and multidrug-resistant tuberculosis, the increase in mobile populations, and co-infection of HIV and tuberculosis. Tuberculosis is still one of the most serious infectious diseases in the world today. One-third of the world's population and about 2 billion people have been infected with Mycobacterium tuberculosis. There are about 10 million new infections every year, and about 3 million people die from it every year. tuberculosis. There are currently 5 million tuberculosis patient...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/53C07H21/00
Inventor 张灵霞吴雪琼
Owner THE 309TH HOSPITAL OF CHINESE PEOPLES LIBERATION ARMY
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