Method for synchronous detection of multiple tubercle bacillus specific secretion antigens

A technology for simultaneous detection of Mycobacterium tuberculosis, applied in the field of medical testing, can solve the problems of low sensitivity and specificity, and achieve the effect of improving sensitivity and improving the positive detection rate.

Inactive Publication Date: 2006-11-22
SHANGHAI PULMONARY HOSPITAL +1
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Problems solved by technology

[0007] The purpose of the present invention is to provide a method for synchronously detecting multiple Mycobacterium tuberculosis-specific secreted antigens for

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  • Method for synchronous detection of multiple tubercle bacillus specific secretion antigens

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Example Embodiment

[0029] Example 1

[0030] 1) Through the analysis of the gene sequence of the specific antigen of Mycobacterium tuberculosis, it is determined that the main secreted antigens of ESAT-6, MPT53, MPT64, MPT63, and MPT70 are the candidate genes.

[0031] Different fusion methods are used for vector construction and expression. The vector uses pET21a, the C-terminal contains 6×His after expression, pProEX-HTa, and the N-terminal contains 6×His after expression. Table 1 shows the results of different fusion methods and vector construction and expression.

[0032] Carrier

[0033] The gene sequence of the above protein to be expressed was cloned into the vector, transfected into E. coli, induced, expressed, and purified. The purified protein was analyzed and quantified by SDS-PAGE, and the pure product of the above antigen was obtained as the immunogen.

[0034] 2) Antibody preparation: prepare polyclonal antibodies with MPT63-MPT70 and ESAT-6-MPT53-MPT64 according to the relev...

Example Embodiment

[0054] Example 2 Serum detection of patients infected with Mycobacterium tuberculosis

[0055] In the same way as in step 6 of Example 1, 30 samples of Mycobacterium tuberculosis infection patient sera and 10 samples of other cases were tested. 24 sera of 30 TB-positive patients were judged to be positive (80%). All 10 TB-negative sera samples were tested. It is negative (cutoff value is 0.261).

[0056] Table 3 shows the results of 30 sera of patients infected with Mycobacterium tuberculosis.

[0057] TB positive

[0058] Using the method of the present invention and the single antigen detection method of known technology, 30 samples of sera from patients with Mycobacterium tuberculosis infection and sera of 30 other cases were tested by two different methods. The test results show that the method of the present invention can simultaneously detect the specificity of 5 species of Mycobacterium The sensitivity of secreting antigens to positive detection of tuberculosis pa...

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Abstract

The related detection method for bacillus tubercle specific secretory antigen comprises: culturing or using bio-engineering method to obtain ESAT-6, MPT53, MPT64, MPT63 and MPT70, preparing opposite antigen, and realizing synchronous detection for five antigens. This invention improves detection sensitivity and can provide effective evaluation for diagnosis.

Description

technical field [0001] The invention belongs to the field of medical testing, and relates to a method for diagnosing Mycobacterium tuberculosis infection and evaluating the curative effect against tuberculosis, in particular to a method for synchronously detecting multiple specific antigens secreted by Mycobacterium tuberculosis. Background technique: [0002] Tuberculosis is still a public health problem that seriously endangers humans and animals, and human tuberculosis is an infectious disease that WHO focuses on controlling. At present, about 1 / 3 of the world's population is infected with tuberculosis, and there are about 20 million tuberculosis patients, with 8 million to 10 million new cases each year. About 2 million people die from tuberculosis every year, more than all other infectious diseases combined. The tuberculosis epidemic in my country is extremely severe, and 1 / 4 of the world's tuberculosis patients are in China. There are currently 500 million people inf...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/543G01N33/569
Inventor 胡忠义丁元生杨华王洁秦莲花
Owner SHANGHAI PULMONARY HOSPITAL
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