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Composition based on tubercle bacillus antigenic polypeptide for diagnosing tuberculosis

A composition and technology for tuberculosis, applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve the problem of not finding the sensitivity and specificity of tuberculosis

Inactive Publication Date: 2009-07-08
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] However, in this field, a satisfactory combination of sensitivity and specificity in the detection of tuberculosis has not been found

Method used

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  • Composition based on tubercle bacillus antigenic polypeptide for diagnosing tuberculosis
  • Composition based on tubercle bacillus antigenic polypeptide for diagnosing tuberculosis
  • Composition based on tubercle bacillus antigenic polypeptide for diagnosing tuberculosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Determination of seven kinds of Mycobacterium tuberculosis specific epitope antigen peptides

[0031] Using the BIOSUN biological analysis software, the epitope distribution of 7 Mycobacterium tuberculosis antigens 38kD, ESAT-6, CFP10, MPT64, Mtb8, Mtb8.4 and Mtb16.3 were analyzed respectively. , the obtained epitope distribution map is shown in Figure 1-7 . According to the epitope distribution map, the specific epitope antigen peptide was finally determined. The coding DNA sequence and amino acid sequence of the specific epitope antigen peptide of Mycobacterium tuberculosis antigen 38kD are respectively SEQ ID NO: 1 and SEQ ID NO: 2; the coding DNA sequence of the specific epitope antigen peptide of Mycobacterium tuberculosis antigen ESAT-6 and the amino acid sequence are respectively SEQ ID NO: 3 and SEQ ID NO: 4; the coding DNA sequence and the amino acid sequence of the specific epitope antigen peptide of Mycobacterium tuberculosis antigen CFP10 are re...

Embodiment 2

[0032] Example 2: Preparation of 7 separate Mycobacterium tuberculosis specific epitope antigen peptides

[0033] 1. Cloning of specific epitope antigen peptide gene

[0034] The upstream and downstream primers were designed and synthesized according to the nucleotide sequence of the specific epitope antigen peptide, and the restriction enzymes used were BamH I and Xho I respectively.

[0035] The primer sequence used to amplify the 38kD specific epitope antigen peptide is as follows:

[0036] 38kD-F: 5'-GGGATCCGCGGCGGGCTGTGGCTCGA-3'

[0037] 38kD-R: 5'-GCTCGAGGCTGGAAATCGTCGCGA-3'

[0038] The primer sequences used to amplify the specific epitope antigen peptide ESAT6 are as follows:

[0039] ESAT6-F: 5'-GGGATCCCATTCCCTCTTGACGA-3'

[0040] ESAT6-R: 5'-GCTCGAGGTTCAGCTCGGTAGCCGT-3'

[0041] The primer sequences used to amplify the specific epitope antigen peptide CFP10 are as follows:

[0042] CFP10-F: 5'-CGGATCCGAAGCAGCCAATAAGCAG-3'

[0043] CFP10-R: 5'-GCTCGAGCGAGGACAGC...

Embodiment 3

[0062] Example 3: Preparation of 38kD-ESAT6-CFP10 Fusion Antigen

[0063] 1. Primer Design

[0064] The upstream and downstream primers were designed according to the sequences of the three antigen genes and the linker. The restriction enzymes used were BamH I and Xho I respectively. All primers were synthesized by Shanghai Handsome Biotechnology Company, and their sequences are as follows:

[0065] 38kD-F: 5'-GGGATCCGCGGCGGGCTGTGGCTCGA-3'

[0066] 38kD-RI: 5'-ACTACCGCCACCAGAGCTGGAAATCGTCGC-3'

[0067] ESAT6-FL: 5'-AGCGGCGGCGGTAGCCATTCCCTCTTGAC-3'

[0068] ESAT6-RL: 5'-AGAGCCACCGCCACTGTTCAGCTCGGTAGC-3'

[0069] CFP10-FL: 5'-CTTCCGGTGGTGGCTCTGAAGCAGCCAATAA-3'

[0070] CFP10-R: 5'-GCTCGAGCGAGGACAGCGCCTGCTG-3'

[0071] 2. Vector Construction

[0072]Using the genomic DNA of Mycobacterium tuberculosis strain H37Rv as a template, gene fragments of 38kD, ESAT-6 and CFP10 specific epitope antigen peptides were respectively amplified. PCR amplification conditions are as follows...

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Abstract

The invention relates to a composition used for serological diagnosis of tuberculosis, which comprises a combination of a special antigen epitope peptide of tubercle bacillus Mtb16.3 with at least one tubercle bacillus special antigen epitope peptide selected from 38kD, ESAT-6, CFP10andMPT64.

Description

technical field [0001] The present invention relates to the field of diagnosis of tuberculosis. More specifically, it relates to the field of diagnosis of tuberculosis using a combination of multiple antigenic polypeptides derived from Mycobacterium tuberculosis. Background of the invention [0002] In recent years, the resurgence of tuberculosis has become a public health and social issue of global concern. According to statistics, there are currently 20 million tuberculosis patients in the world, 10 million new cases, and 3 million deaths per year, surpassing the sum of other infectious diseases and becoming the number one killer of infectious diseases. [0003] Tuberculosis exhibits all the main features of a global epidemic and is spreading worldwide. About 1 / 3 of the people on the planet are infected with Mycobacterium tuberculosis, resulting in 8 million cases of active tuberculosis and 3 million deaths each year. The emergence of multidrug-resistant strains of Myco...

Claims

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Application Information

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IPC IPC(8): G01N33/569
Inventor 冯晓燕张贺秋陈坤宋晓国王国华
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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