Tuberculosis diagnostic composition and application thereof

A diagnostic composition and composition technology, applied in the field of tuberculosis detection, can solve the problems of complex composition, cumbersome operation, and high cost of the diagnostic reagent antigen peptide library, and achieve the effects of simplifying the detection design and operation process, improving sensitivity, and reducing diagnostic costs

Active Publication Date: 2012-07-25
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The sensitivity of existing diagnostic reagents needs to be further improved. The antigen peptide library of diagnostic reagents based on ELISPOT technology is complex in composition and cumbersome in operation, and

Method used

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  • Tuberculosis diagnostic composition and application thereof
  • Tuberculosis diagnostic composition and application thereof
  • Tuberculosis diagnostic composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Composition of embodiment 1

[0034] A composition for diagnosing tuberculosis, consisting of antigen ESAT-6-derived polypeptides, antigen CFP-10-derived polypeptides and antigen Rv3615c-derived polypeptides;

[0035] The amino acid sequence of the antigen ESAT-6 is shown in SEQ ID NO.1.

[0036] The amino acid sequence of the antigen CFP-10 is shown in SEQ ID NO.2.

[0037] The amino acid sequence of the antigen Rv3615c is shown in SEQ ID NO.3.

[0038] The amino acid sequence of the ESAT-6 derivative polypeptide is shown in SEQ ID NO.4 to SEQ ID NO.20.

[0039] The amino acid sequence of the CFP-10 derivative polypeptide is shown in SEQ ID NO.21 to SEQ ID NO.38.

[0040] The amino acid sequence of the Rv3615c derivative polypeptide is shown in SEQ ID NO.39 to SEQ ID NO.57. Embodiment 2 antigen is applied to the development of detection kit

Embodiment 2

[0040] The amino acid sequence of the Rv3615c derivative polypeptide is shown in SEQ ID NO.39 to SEQ ID NO.57. Embodiment 2 antigen is applied to the development of detection kit

[0041] The antigen protected by the invention can be used for detection of mycobacterium tuberculosis infection, and can form a kit for clinical or laboratory diagnosis.

[0042] The test kit contains:

[0043] 1. The mixture described in Example 1;

[0044] 2. Phytohemagglutinin (PHA) solution or other non-specific positive stimulators of T cells;

[0045] 3. ELISPOT 96-well PVDF membrane plate;

[0046] 4. Human γ-interferon monoclonal antibody (primary antibody), biotin-labeled human γ-interferon monoclonal detection antibody (secondary antibody), horseradish peroxidase-labeled streptavidin and horseradish peroxidase Oxidase reaction substrate;

[0047] 5. Other reagents and consumables required for ELISPOT detection.

Embodiment 3

[0048] The antigen of Example 3 is applied to the clinical detection of Mycobacterium tuberculosis infection.

[0049] A. Peripheral blood lymphocyte separation:

[0050] The volunteer case screening criteria involved in the present invention are:

[0051] 1) The clinical attending doctor diagnosed pulmonary tuberculosis according to the clinical manifestations of the volunteers;

[0052] 2) Imaging studies show shadows in the lungs;

[0053] 3) Mycobacterium tuberculosis culture is positive or negative.

[0054] The screening criteria for healthy volunteers are:

[0055] 1) No clinical symptoms of tuberculosis;

[0056] 2) No other diseases or infections.

[0057] The selected tuberculosis cases and healthy volunteers were between 18 and 60 years old, and their sex was randomly selected according to 1:1.

[0058] The T lymphocytes used in the present invention are obtained from the venous peripheral blood of an individual. After the selected individuals pass the physic...

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Abstract

The invention relates to a tuberculosis diagnostic composition and application of the tuberculosis diagnostic composition, in particular provides the tuberculosis diagnostic composition which consists of optional two or three of antigen ESAT-6 derived polypeptide, antigen CFP-10 derived polypeptide and antigen Rv3615c derived polypeptide. The invention also provides application of the tuberculosis diagnostic composition in a specific T cellular immune reaction caused by the detection of mycobacterium tuberculosis infection in vitro and application of the preparation of a tuberculosis diagnostic agent. Compared with the existing diagnostic technology, the diagnostic cost is reduced, the detection design and the operation process are simplified, and the detection sensitivity is obviously improved.

Description

technical field [0001] The invention relates to a tuberculosis diagnostic composition and its application, belonging to the technical field of tuberculosis detection. Background technique [0002] Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis infection. According to estimates from the World Health Organization, one-third of the world's population is infected by Mycobacterium tuberculosis. There are more than nine million new cases of active infection and one million deaths each year. example above. In recent years, the incidence of tuberculosis in my country has remained high, with more than one million new cases and nearly 200,000 deaths each year, which has seriously harmed the health of our citizens, social stability and economic development. Tuberculosis is a respiratory infectious disease that seriously endangers the health of the people and is listed as one of the major infectious diseases in my country. Although the rising momentum of th...

Claims

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Application Information

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IPC IPC(8): G01N33/68C12N15/31C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A61K39/04A61K48/00A61P31/06
Inventor 高福谭曙光范盘生张宁万康林
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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