97 results about "Mycobacterium tuberculosis antigens" patented technology

The present invention relates to fusion proteins of Mycobacterium tuberclosis antigens. In particular, it relates to two fusion proteins, each of which contains three individual M. tuberculosis antigens, and a fusion protein of two M. tuberculosis antigens, their coding sequences, and methods for their use in the treatment and prevention of tuberculosis.

The present invention relates to MHC-peptide complexes and uses thereof in the diagnosis of, treatment of or vaccination against a disease in an individual. More specifically the invention discloses MHC complexes comprising Mycobacterium tuberculosis antigenic peptides and uses there of.

The present invention relates to fusion proteins containing at least two Mycobacterium species antigens. In particular, it relates to nucleic acids encoding fusion proteins that include two or more individual M. tuberculosis antigens, which increase serological sensitivity of sera from individuals infected with tuberculosis, and methods for their use in the diagnosis, treatment, and prevention of tuberculosis infection.

The present invention relates to a method to prepare a reagent kit to quickly diagnose mycobacterium tuberculosis antigens jointly packaged by 38KD and 16KD antigens and a test method. The blood antigen test paper of the present invention jointly packages 38 KD and 16KD of TB antigens into a lower part of a NC film to form a test line (T). Goat anti-mouse IgG is packaged into an upper part of the NC film to form a quality control line (C). In addition, anti-human IgG (mouse anti-human IgG, goat anti-human IgG or SPA) marks nanometer coloring particles (colloidal gold particles or sol particles or nanometer beads) to prepare a labeling pad. The NC film, the labeling pad, a sampling pad and a sample adsorption pad are glued and assembled onto a plastic board to form the test paper. During test, few tested sample is added onto the NC film of the test paper. And then, sample diluents are added through a sampling end. After sampling, it is necessary to observe result of immunoreaction, thus realizing quick auxiliary TB antigen diagnosis.

A method for detection of mycobacterium tuberculosis antigens in biological fluids provides immunoassay methods, diagnostic kits, and an immunochromatoraphic assay device for detection of Mycobacterium tuberculosis antigens in biological specimens, preferably body fluids and tissues. The preferred body fluids are blood, serum, plasma, urine, pulmonary fluid, sputum, cerebrospinal fluid, and the preferred tissue is the lung biopsy specimen. The immunoassays require two primary antibodies against RD1, RD2, or RD3 of Mycobacterium tuberculosis. At least one of the primary antibodies is attached to a solid carrier. Optional, a second antibody against an animal species producing one of the primary antibodies can be added. Either the other primary antibody or the secondary antibody is labeled with a detection agent, which can be an enzymatic marker, a fluorescent or luminescent agent, a radio active label or a color particle. The biological specimens may be used directly, concentrated or diluted for the immunoassays.

The present invention is based on the identification and characterization of a number of novel M. tuberculosis derived proteins and protein fragments. The invention is directed to the polypeptides and immunologically active fragments thereof, the genes encoding them, immunological compositions such as vaccines and skin test reagents containing the polypeptides.

The invention discloses a tuberculosis gene vaccine based on T cell epitopes, wherein a full-length gene, embedded with four T cell epitope polypeptide genes which come from mycobacterium tuberculosis antigen, of mycobacterium tuberculosis heat shock protein is inserted into a vector. The invention also discloses a method for preparing the vaccine, which comprises the following steps: four T cell epitope genes, namely EAST-6189-228, Ag85A369-405, CFP10162-207 and Ag85B420-459 which come from the mycobacterium tuberculosis antigen are inserted into an HSP65 full-length gene. The invention also discloses application of an ECANS tuberculosis gene vaccine. Through the intramuscular injection of the gene vaccine into an immune mouse, the experiment proves that the vaccine can induce a specific antibody which aims at a plurality of tuberculosis antigens to response, can induce stronger tuberculosis specific killing response, can induce Th1 immune response at the same time, secrete high-level IFN gamma, and is a good vaccine for preventing and treating tuberculosis.

The present invention relates to fusion proteins containing at least two Mycobacterium tuberculosis antigens. In particular, it relates to bi-fusion proteins which contain two individual M. tuberculosis antigens, tri-fusion proteins which contain three M. tuberculosis antigens, tetra-fusion proteins which contain four M. tuberculosis antigens, and penta-fusion proteins which contain five M. tuberculosis antigens, and methods for their use in the diagnosis, treatment and prevention of tuberculosis infection.

An iontophoresis device includes an active electrode assembly including an active electrode element and at least one active agent reservoir. Active agents include one or more polypeptides, or fusion polypeptides, having antigenic determinants derived from or related to M. tuberculosis antigens and suitable for detection of tuberculosis.

The present invention relates to a method for diagnosing latent tuberculosis infection in a subject comprising the step i) consisting of incubating a biological sample obtained from the subject with at least one Mycobacterium tuberculosis antigen, and thereafter a step ii) consisting of quantifying in said biological sample the secretion of at least one cytokine selected from the group consisting of IL-2, IL-15, IP-10 and MIG.

The present invention is based on the identification and characterization of a number of novel M. tuberculosis derived proteins and protein fragments. The invention is directed to the polypeptides and immunologically active fragments thereof, the genes encoding them, immunological compositions such as diagnostic reagents containing the polypeptides.

The invention is related to an immunogenic composition, vaccine or pharmaceutical composition for preventing, boosting or treating infection caused by a species of the tuberculosis complex (M. tuberculosis, M. bovis, M. africanum, M. microti). The immunogenic composition, vaccine or pharmaceutical composition comprise a fusion polypeptide, which comprises one or more starvation antigens from M. tuberculosis, the units of the fusion polypeptide being M. tuberculosis antigens. Further, the invention is related to the use of a vaccine comprising a fusion polypeptide sequence or nucleic acid sequence of the invention given at the same time as BCG, either mixed with BCG or administered separately at different sites or routes for preparing said immunogenic composition, vaccine, or pharmaceutical composition.

One mycobacterium tuberculosis antigen detective method in tuberculosis bacilli solution, including the following steps: (a) Choice and prepare TB-specific antigen that can be detect by the antigen-antibody binding reactions in various body fluids; (b) Prepare of confrontation and strains of TB-specific antigen monoclonal antibody; (C) Prepare TB detection reagent kit of specific antigen secreted; (d) Use the above-mentioned antibody and antigen detection kit for tuberculosis. Take these steps and reach high rate of the test and correct testing purposes.

The invention discloses a mycobacterium tuberculosis fusion protein and an application thereof in induction of peripheral blood mononuclear cells (PBMCs) to generate cytokines. The fusion protein includes three proteins PPE41, ESAT-6 and PE25, and the proteins are connected through connecting peptides. Compared with present stimulants, the fusion protein provided by the invention has the advantages of efficient effect, strong sensitivity, high specificity and good stimulation effect. The fusion protein stimulates the PBMCs to generate a large amount of mycobacterium tuberculosis antigen specific IFN-gamma, IL-2, TNF-alpha and other tuberculosis related factors, and the above stimulation induction reaction is free from BCG vaccine interference. The fusion protein can effectively improve the tuberculosis detection rate and is of positive significance to control the tuberculosis. The fusion protein can be applied in researches of the tuberculosis pathopoiesis and immunoprophylaxis mechanisms and control of the tuberculosis as a stimulant.

The invention discloses construction and application of a mycobacterium tuberculosis fusion protein, and relates to construction of fusion proteins. Specific antigens of bacteria in a dormant period are omitted in the prior mycobacterium tuberculosis fusion protein; and the mycobacterium tuberculosis fusion protein provided by the invention contains a protective antigen HspX, a segment of amino-peptides from 190 position to 198 position of Mpt64 of CD8<+>T-cell epitopes and a mycobacterium tuberculosis antigen 85 complex Ag85B. The protective antigens in the dormant period are screened; the protective antigens in the dormant period and antigens in the exponential phase are combined to construct a vaccine for attempting to create immunoreaction aiming at bacterial communities of the mycobacterium tuberculosis in different growth states so as to effectively eliminate or monitor multiple kinds of mycobacterium tuberculosis including latent infection and prevent the occurrence of tuberculosis of adults.

The present invention relates to applications of Mycobacterium tuberculosis antigen protein Rv0446c and a T-cell epitope peptide thereof in preparation of tuberculosis detection reagents, vaccines and medicines, wherein the amino acid sequences of the antigen protein Rv0446c and the T-cell epitope peptide thereof are respectively represented by SEQ ID NO:1-5. According to the present invention, the Mycobacterium tuberculosis antigen protein Rv0446c and the T-cell epitope peptide thereof are used as the stimulants for the specific T cell and B cell immune response caused by Mycobacterium tuberculosis infection, and the false positive caused by the impure antigen can be reduced compared with the use of the complete antigen in the prior art; and the detection reagents prepared from the antigen protein Rv0446c and the T-cell epitope peptide thereof can be widely used for assisted diagnosis of tuberculosis, epidemiological surveillance and other related fields, and the tuberculosis vaccines and the anti-tuberculosis drugs prepared from the antigen protein Rv0446c and the T-cell epitope peptide thereof can be used for prevention and treatment of tuberculosis.

The invention discloses an oligonucleotides aptamer of targeted mycobacterium tuberculosis Ag85B, a preparation method and application thereof. The oligonucleotides aptamer has nucleotide sequences shown as AP-1 to AP-13. The preparation method of the oligonucleotides aptamer comprises steps as follows: (1) constructing a random single-stranded DNA (ssDNA) library and preparing a primer; (2) preparing the random single-stranded DNA (ssDNA) library by PCR amplification; (3) carrying out SELEX screening; (4) detecting the appetency; (5) cloning and sequencing DNA. The oligonucleotides aptamer of targeted mycobacterium tuberculosis Ag85B is applied to detect the antigen level of the mycobacterium tuberculosis Ag85B in human serum, can replace antibodies, improves the sensitivity and the specificity of the tuberculosis antigen detection in specimens of body fluid (such as serum, hydrothorax, cerebrospinal fluid ascites and the like), thereby improving the accuracy rate of tuberculosis diagnosis.

The invention provides a combined antigen comprising three mycobacterium tuberculosis antigens and an application of the combined antigen to preparation of a reagent for diagnosing active tuberculosis. The combined antigen comprises Rv3871, Rv3876 and Rv3879. Corresponding antibody detection methods are established by the aid of the three antigens and used for detecting corresponding antibody levels in biological samples, and the three detected antibody levels are particularly higher in sputum smear negative/sputum culture positive tuberculosis patients. A pulmonary tuberculosis diagnosing method is established by the aid of the combined antigen, and a positive sample is defined in such a manner that antibody levels corresponding to two or more antigens reach a positive judgment value. Verification of a lot of clinical samples indicates that the diagnosing method has the high sensitivity and the high specificity for sputum smear positive/sputum culture positive, sputum smear negative/sputum culture positive and sputum smear negative/sputum culture negative tuberculosis patients.

The invention aims to solve the technical problem that an antibody of mycobacterium tuberculosis in blood serum is difficult to rapidly and more accurately detect in the field. The technical scheme of the invention is to provide a detection reagent and rapid detection method for detecting an antibody of mycobacterium tuberculosis in blood serum. The detection reagent of the antibody of the mycobacterium tuberculosis in the blood serum comprises three types of mixed mycobacterium tuberculosis antigen proteins, namely 38kDA antigen protein, CFP10 antigen protein and MPT64 antigen protein. The detection method of the antibody of the mycobacterium tuberculosis in the blood serum comprises the following steps of: contacting in-vitro blood serum with the reagent containing the 38kDA antigen protein, the CFP10 antigen protein and the MPT64 antigen protein of the mycobacterium tuberculosis; and judging whether the in-vitro blood serum contains the mycobacterium tuberculosis according to the result of a process for detecting whether an antigen-antibody reaction exists. The reagent provided by the invention has the characteristics of fastness, sensitivity, specialty, simplicity, convenience, and the like; the reagent can be used for rapidly sieving the antibody of the mycobacterium tuberculosis in the in-vitro blood serum, is applicable to each grade of laboratories, and has a very good application prospect.

The invention relates to 12 mycobacterium tuberculosis antigen polypeptides, and antigen epitope polypeptides having polypeptide sequences having at least 90% of the same as each other; the antigen polypeptides or the antigen epitope polypeptides are matched with an immunologic adjuvant to obtain mycobacterium tuberculosis vaccines; the antigen polypeptides or the antigen epitope polypeptides are used as antigens, and monoclonal antibodies are prepared and used for detecting whether the sample contains mycobacterium tuberculosis; nucleic acid sequences of the 12 mycobacterium tuberculosis antigen polypeptides and polynucleotides having the nucleic acid sequences having at least 90% of the same as each other are encoded; gene vaccines containing the nucleic acids and the polynucleotides are prepared. Beneficial effects comprise that the polypeptides can be used for recombined vaccine polypeptides, or be used for development of antigen diagnostic reagents, so as to be beneficial for detection and diagnosis of pulmonary tuberculosis.

The invention relates to one or more types of recombinant fusion proteins formed by hepatitis B core proteins and mycobacterium tuberculosis antigens or antigen fragments. The tuberculosis antigens or antigen fragments are inserted in the same or different permission sites of hepatitis B core proteins in a mode of single antigen or antigen fragment or various types of antigens or antigen fragments which are combined together. The recombinant fusion proteins may comprise a plurality of cofactors. In addition, the invention also relates to the type of the fusion of the tuberculosis antigen or antigen fragments and cofactors and hepatitis B core proteins, nucleic acid and amino acid sequences of coded corresponding recombinant fusion proteins, a method for preparing the recombinant fusion proteins and application of the recombinant fusion proteins in the preparation of medicaments and vaccines for preventing and/or treating tuberculosis.

The invention provides an antigen composition used for immunodetection of a tuberculosis infected cell. The antigen composition comprises a mycobacterium tuberculosis Rv3873 antigen and/or antigen epitope peptides derived from the antigen. In an optimized embodiment, the antigen composition comprises all the antigen epitope peptides contained in the mycobacterium tuberculosis Rv3873 antigen. The invention also provides an application of the antigen composition to preparation of detection reagents used for detecting specific T cell immune response caused by tuberculosis infection in vitro. After lymphocytes of human or animals are stimulated by the antigen composition, cell factors secreted by a tuberculosis specific T cell are detected.