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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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 the defects of low sensitivity and specificity in the detection of Mycobacterium tuberculosis and the evaluation of anti-tuberculosis curative effect at present

Method used

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

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1) Through the analysis of the gene sequence of the antigen with strong specificity to Mycobacterium tuberculosis, several major secreted antigens of ESAT-6, MPT53, MPT64, MPT63 and MPT70 are determined as candidate genes.

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

[0032] carrier

expression product

deduced product sequence

The expression level

pET21a

pET21a

pET21a

pProEX-HTa

pProEX-HTa

pProEX-HTa

MPT63-6×His

MPT70-6×His

MPT63-MPT70-6×His

6×His-MPT64

6×His-ESAT-6-MPT53

6×His-ESAT-6-MPT53-MPT64

SEQ1

SEQ2

SEQ3

SEQ4

SEQ5

SEQ6

Low expression level, unable to purify

Low expr...

Embodiment 2

[0054] Example 2 Serum Detection of Mycobacterium tuberculosis Infected Patients

[0055] With the step 6 method of embodiment 1, 30 parts of Mycobacterium tuberculosis infection patient's serum and 10 other case serums are tested, 24 routine test results of 30 routine TB positive patient's serums are judged as positive (80%), 10 routine TB negative serum samples all Negative (cutoff value 0.261).

[0056] Table 3 shows the test results of 30 tubercle bacillus infected patients' sera.

[0057] TB positive

serum sample

OD

value

TB positive

serum sample

OD

value

TB positive

serum sample

OD

value

TB negative

serum sample

OD

value

1

0.041

11

1.221

21

0.332

1

0.075

2

0.756

12

0.562

22

0.246

2

0.039

3

0.864

13

0.769

23

0.842

3

0.046

4

1.256 ...

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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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