Antigen composition used for immunodetection of tuberculosis infected cell and application thereof
A technology for infecting cells and immune detection, applied in the field of tuberculosis detection, can solve the problems such as the need to improve the sensitivity and specificity, high cost, complex composition of antigen components, etc., to achieve improved detection sensitivity, low cost, and suitable for large-scale commercial production. Effect
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Embodiment 1
[0044] The composition of embodiment 1 epitope peptide composition
[0045] The composition for tuberculosis detection consists of epitope peptide derived from CFP-10 antigen, ESAT-6 antigen and Rv3873 antigen.
[0046] The amino acid sequence of the CFP-10 antigen is shown in SEQ ID NO.1.
[0047] The amino acid sequence of the ESAT-6 antigen is shown in SEQ ID NO.2.
[0048] The amino acid sequence of the Rv3873 antigen is shown in SEQ ID NO.3.
[0049] The amino acid sequence of the epitope peptide derived from the CFP-10 antigen is selected from SEQ ID NO.4-59.
[0050] The amino acid sequence of the epitope peptide derived from ESAT-6 antigen is selected from SEQ ID NO.60-122.
[0051] The amino acid sequence of the epitope peptide derived from the Rv3873 antigen is selected from SEQ ID NO.123-180.
Embodiment 2
[0052] Example 2 Antigen epitopes are applied to the development of detection kits
[0053] The antigen epitope peptide composition of the present invention can be used for the detection of tuberculosis infection, and can form a kit for clinical or laboratory diagnosis.
[0054] The test kit includes:
[0055] 1) the epitope peptide composition described in Example 1;
[0056] 2) mouse IgG monoclonal antibody (primary antibody) against human or animal IFN-γ, which is fixed on the reaction plate;
[0057] 3) Enzyme-labeled reagent: another mouse IgG monoclonal antibody (secondary antibody) labeled with horseradish peroxidase against different epitopes of human or animal IFN-γ;
[0058] 4) Calibrator: positive raw material containing IFN-γ;
[0059] 5) Culture tubes: the test culture tubes contain artificially synthesized epitope polypeptides derived from various antigens, the positive control culture tubes contain tuberculosis non-specific stimulating antigens, and the backg...
Embodiment 3
[0061] Example 3 Antigen epitopes are applied to clinical detection of tuberculosis infection
[0062] A. Isolation of peripheral blood lymphocytes:
[0063] The screening criteria of volunteer cases involved in the present invention are:
[0064] According to the national standard (WS288-2008) of the "Diagnostic Criteria for Pulmonary Tuberculosis" issued by the Ministry of Health, patients with clinical manifestations, symptoms, signs and chest imaging examinations diagnosed as pulmonary tuberculosis, among which bacteriologically positive or negative tuberculosis patients refer to patients clinically diagnosed as tuberculosis Among them, tuberculosis patients with positive or negative bacteriological examination (microscopic examination of sputum smear acid-fast staining and / or bacterial solid culture on Roche solid medium); and extrapulmonary tuberculosis (such as bone tuberculosis, renal tuberculosis, intestinal tuberculosis, lymphatic tuberculosis) etc.) patients.
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