Tuberculosis diagnostic composition and application thereof
A diagnostic composition and composition technology, applied in the field of tuberculosis detection, can solve the problems of cumbersome operation, high cost, complex composition of diagnostic reagent antigen peptide library, etc., and achieve the goal of improving sensitivity, reducing diagnostic cost, and simplifying detection design and operation process Effect
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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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