190 results about "Active tuberculosis" patented technology

The invention belongs to the field of biomedicine examination, and particularly relates to a kit and a method for detecting mycobacterium tuberculosis infection and application. The invention discloses a novel mycobacterium tuberculosis detection reagent by screening specific T cell epitope of mycobacterium tuberculosis, wherein the reagent contains polypeptide or analog thereof represented by SEQ ID No.1-10. The method detects cell factors released from T cells by using single or more SEQ ID No.1-10 polypeptides to contact the T cells of mycobacterium tuberculosis infected individuals. The method can effectively detect active tuberculosis or latent tuberculosis infection, and is free from disturbance of Bacilli Calmette Guerin (BCG) inoculation vaccines. The invention also discloses a diagnostic kit and other application based on the polypeptide and the method. Compared with the gamma interferon release experiments in the prior art, the method can obviously improve the detection rate without reducing the specificity and has high clinical application value.

The invention discloses a kit for assisted diagnosis of tuberculosis, which comprises a specific antibody composition, wherein the specific antibody composition comprises the following five antibodies: (1) IFN-gamma antibody, (2) TNF-alpha antibody, (3) IL-2 antibody, (4) MIG antibody and (5) IP-10 antibody. The kit disclosed by the invention can distinguish a Mycobacterium tuberculosis infected person from a BCG vaccinee. Compared with the ELISPOT tuberculosis diagnosis method, the tuberculosis diagnosis kit based on multi-molecular marker detection has obviously enhanced sensitivity to active tuberculosis (from 72% to 89%), and the positive rate for normal healthy persons is obviously reduced (from 27% to 16%).

The invention belongs to the biomedical detection field, in particular relates to a reagent and method used for detecting activity tuberculosis and latent tuberculosis infection; based on the genomic principle, the invention discloses a novel detection reagent for mycobacterium tuberculosis, containing protein or polypeptide which is represented by SEQ ID1-2, 4-5, 8-28; the method uses one or a plurality of SEQ ID 1-28 protein or polypeptide to contact T cells of a mycobacterium tuberculosis host, and detects cytokine released from the T cells; the method can detect the tuberculosis and latent infection effectively and is not interfered by BCG vaccine at the same time; the invention also discloses a diagnostic reagent kit and other application based on the protein or polypeptide and the method; compared with the T-SPOT of the prior art, the invention can improve detectable rate obviously under the condition that the specificity is not reduced; the reagent kit has cheap price, and the cost is 1/5 to 1/10 of that of the T-SPOT reagent, thus being beneficial to being popularized in the developing countries and poor areas.

Improved methods for detecting active tuberculosis are disclosed. A method comprises enriching at least one M. tuberculosis-specific biomolecule from a sample by contacting the sample with a nanoporous film; and detecting the presence of the M. tuberculosis-specific biomolecule or fragment(s) thereof. The method may further comprise digesting the enriched M. tuberculosis-specific biomolecule with an enzyme to produce a digestion product comprising at least one fragment of the M. tuberculosis-specific biomolecule. Improved sensitivity and speed achieved.

The invention discloses a kit for detecting active tuberculosis based on antigen-specific TNF-alpha-ELISA (enzyme linked immunosorbent assay) and an application thereof. The kit comprises an ELISA plate coating a TNF-alpha antibody, a TNF-alpha protein standard, protein liquid of ESAT-6 and CFP-10, a biotin-marked TNF-alpha antibody and avidin-marked horseradish peroxidase. By adopting the kit for detecting active tuberculosis based on TNF-alpha-ELISA, a method for distinguishing active tuberculosis infection from latent tuberculosis infection is established; the TNF-alpha released by the peripheral blood mononuclear cell under the stimulus of tuberculosis-specific antigens ESAT-6 and CFP-10 and the background TNF-alpha released by the peripheral blood mononuclear cell without stimulus are quantitatively detected through the ELISA technology, and the value of the antigen-specific TNF-alpha is obtained by subtracting the two so as to perform direct diagnosis on the active tuberculosis.

The invention relates to an establishing method of a serum diagnosing model of active tuberculosis, which has the following steps: step 1 is a serum source taking and preparing step which comprises an experiment group and a control group, the venous blood of a checked person is collected, serum and the rest are separately stored after serum separation at the temperature of -80 DEG C in a refrigerator for spare use; step 2 is the step of the mass spectrometric analysis of the serum and comprises sample treatment, protein plate check, spectrometric analysis and data acquisition; step 3 is the step of statistic analysis and the difference protein screening; step 4 is the step of establishing the diagnosing model, and Biomarker Pattern 5.0 software is used for establishing the model of diagnosing active tuberculosis. When the model is used for identifying the active tuberculosis and the control group, the sensitivity is 96.9 percent, the specificity is 97.8 percent, the positive predicating value and the negative predicating value are 97.7 percent and 97.1 percent, and the total accurate rate is 97.3 percent.

The present invention describes compositions for both diagnostic and therapeutic applications. In one embodiment, the present invention contemplates a method of identifying an active M. tuberculosis infection. In another embodiment, the present invention contemplates a method of monitoring a M. tuberculosis infection. In yet another embodiment, the present invention contemplates a method of monitoring a patient's response to treatment for an active M. tuberculosis infection. In a further embodiment, the present invention contemplates a method of monitoring a patient's response to treatment for an active M. tuberculosis infection.

The present invention relates to a method of diagnosing and monitoring various distinct presentations of tuberculosis: active tuberculosis disease, latent tuberculosis infection and recent tuberculosis infection. The rapid immune assay is based on the evaluation of the frequency of Interferon (IFN) gamma-producing antigen-specific T lymphocytes responding to selected peptide sequences from Mycobacterium tuberculosis, selected for their immunogenicity. The invention concerns also immunogenic and vaccine compositions based on these specific peptide sequences.

The invention provides a diagnostic kit for detecting active pulmonary tuberculosis. The kit consists of an RNA extract buffer solution, a specific serum miRNA composition for the active pulmonary tuberculosis, internal reference reverse transcription primers, polymerase chain reaction (PCR) primers and a fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) reaction solution, wherein the specific serum miRNA composition for the active pulmonary tuberculosis consists of four differential expression serum miRNAs, namely hsa-miR-29c, hsa-miR-22, hsa-miR-320b and hsa-miR-101. By using the specific serum miRNAs composition for the active pulmonary tuberculosis to detect the active pulmonary tuberculosis on the serum miRNAs level, the sensitivity is 90.2 percent, and the specificity is 75.0 percent; and the kit has early-stage diagnostic value for the active pulmonary tuberculosis, and can realize early warning and early diagnosis of the active pulmonary tuberculosis.

The invention provides a method of detecting surrogate markers for active tuberculosis in a serum sample. The surrogate markers are selected from serum mycolic acid antigen, serum anti-mycolic acid antibodies or both. The method includes the steps of combining the serum sample with a labelled monoclonal immunoglobulin antibody or fragment thereof to mycolic acids to produce a combined serum sample, the antibody or fragment thereof not substantially cross-reacting with cholesterol and the label being selected so that binding of the labelled antibody to immobilized mycolic acid antigen of mycobacterial origin produces a detectable signal and combining a blank sample with the labelled monoclonal immunoglobulin antibody or fragment thereof to mycolic acid to produce a combined blank sample. The method includes exposing both samples to immobilised mycolic acid antigen of mycobacterial origin or a synthetic analogue or analogues thereof so that the labelled immunoglobulin antibodies or fragments thereof in each sample bind to the immobilised antigen to produce detectable signals. If the surrogate markers are present, the signal produced by the blank sample will be stronger than that produced by the serum sample because of inhibition of binding of the labelled antibody in the serum sample arising from prior binding of the labelled antibody with the mycolic acid antigen in the serum sample or by competitive binding of serum anti-mycolic acid antibodies in the serum sample to the immobilised mycolic acid antigen or both.

The invention relates to the technical field of biomarkers, in particular to a biomarker of active pulmonary tuberculosis. The biomarker of active pulmonary tuberculosis is selected from at least one of the following six circRNAs: hsa_circ_0005836, hsa_circ_0009128, hsa_circ_0003519, hsa_circ_0023956, hsa_circ_0078768, hsa_circ_0088452. Specifically, hsa_circ_0005836 has the most obvious down-regulated expression. The biomarker of active pulmonary tuberculosis can be used as a therapeutic target, and is beneficial to development of new drugs.

The invention discloses a miRNA marker for assistant diagnosis of tuberculosis and application thereof. The invention requires the protection of the application of a substance for detecting miRNAs inserum exosomes in the preparation of products used for diagnosing or assisting the diagnosis of tuberculosis patients or for diagnosing or assisting the diagnosis of active tuberculosis patients, wherein each miRNA refers to one or a combination of the following five kinds of miRNAs: miR-28-3p, miR-193b-5p, miR-370-3p, miR-1246 and miR-2110. The invention also requires the application of the substance for detecting miRNAs in serum exosomes in the preparation of products used for diagnosing or assisting the diagnosis of patients with latent tuberculosis infections, wherein each miRNA refers toone or a combination of the following three kinds of miRNAs: let-7e-5p, let-7d-5p and miR-140-5p. A foundation is laid for further development of the early rapid diagnosis technology and method for tuberculosis.

The invention discloses application of lncRNAs as active tuberculosis specific markers. Due to tissue specificity and spatial-temporal specificity of lncRNAs, different tissues are different in lncRNAs expression quantity, and the same tissue or organ varies in lncRNAs expression quantity in different growth stages. Two lncRNAs serving as the active tuberculosis specific markers are disclosed forthe first time, a whole blood sample which can be acquired easily, directly and clinically is combined with designed primer pairs to verify lncRNAs differential expression through RT-qPCR, and quickness, simplicity, time saving, high detection sensitivity and low sample consumption are realized. The method is quite suitable for active tuberculosis diagnosis by lncRNAs in differential expression, and important significances to researching of tuberculosis propagation control and reduction of the tuberculosis occurrence rate are achieved.

The invention discloses a biomarker for diagnosing mycobacterium tuberculosis infection. The biomarker comprises IL-2 and IL-10. The invention proves that cell factors IL-2 and IL-10 can serve as markers for tuberculosis infection diagnosis for the first time, the tuberculosis infection can be diagnosed by measuring the concentration of the IL-2 and the IL-10 after peripheral blood monouclear cells are stimulated by tuberculosis antigen, and active tuberculosis and latent tuberculosis infection are further distinguished. A tuberculosis infection blood detection reagent and a kit which are prepared on the basis of the two cell factors provide new technological basis for quick diagnosis of the active tuberculosis, have the advantages of high sensitivity, high specificity and the like, are quick in operation, reasonable and practical, and can obviously improve the diagnosis efficiency of the active tuberculosis.

The invention provides an MS4A6A gene application, and relates to the preparation of products to distinguish latent tuberculosis infection and active tuberculosis. The preferred products include the products utilizing real-time quantitative PCR or gene chip detection to distinguish the latent tuberculosis infection and the active tuberculosis. According to the experimental results, the expression of the MS4A6A gene is obviously higher in the blood of tuberculosis patients than in healthy people or latent infection crowds, and therefore, the MS4A6A gene can serve as a special marking gene for the diagnosis of tuberculosis, so as to allow the tuberculosis diagnosis to be more accurate and quicker.

The invention relates to a sample containing a tuberculosis serum characterized protein and a preparation method thereof. The sample comprises the serum of an untreated patient suffering from active tuberculosis, and the serum protein of a patient suffering from lung cancer, a patient suffering from pneumonia, a patient suffering from chronic obstructive pulmonary disease or a healthy person who is ensured to be normal by physical examination, wherein the serum protein comprises three up-regulated proteins and one down-regulated protein; the proteins exit independently or any or all of the proteins are mixed together; and the sample is suitable to be detected by using mass spectrum. The sample provides a basis for further discovering new active tuberculosis biomarkers. By using the sample, the detection of the active tuberculosis is superior to any single detection method adopted currently; a non-invasive technology for early discovery and early treatment of the active tuberculosis is provided; and therefore, a new method for reducing the infectious rate of the active tuberculosis and improving the recovery rate of the tuberculosis is provided.

The present invention provides an improved and simplified assay for use in diagnosing active Tuberculosis infections. The present assay is carried out using a sample of whole blood, does not require separating the blood into components, such as the isolation of peripheral blood mononucleocytes (PBMC), and is carried out using only cell-surface staining of T-cells.

The invention discloses application of PRDM1 as a marker for preparing a product for diagnosing active tuberculosis. Fluorescence quantitative PCR is adopted for detecting expression quantity of a PRDM1 gene in a tuberculosis patient PBMCs. A result shows that the relative expression quantity of PRDM1 in smear positive tuberculosis patients, smear negative tuberculosis patients and extrapulmonarytubereulosis patients PBMCs is remarkably lowered that that of latent tuberculosis infected persons and healthy control. A subject working characteristic curve analysis result shows that PRDM1 can distinguish active tuberculosis and latent tuberculosis infection, and becomes the diagnosis target for diagnosing the active tuberculosis.

The invention discloses a co-expression system and construction method of polyvalent bacteriophage lyase genes, a live vaccine of a carrying system and preparation and application of the live vaccine. Eukaryotic expression plasmids are used as an expression vector, and LysinA, LysinB and Holin gene segments are directionally inserted into the plasmids to simultaneously express the co-expression system of the bacteriophage lyase genes of three kinds of targeted mycobacterium tuberculosis. Mycobacterium smegmatis with good targeting property of macrophages or genetically-modified recombinant BCG is used as a live vector, the co-expression system simultaneously carrying three kinds of genes is electrically transformed into the mycobacterium smegmatis or the genetically-modified recombinant BCG, and then the recombinant therapeutic tuberculosis live vaccine is obtained through expansion in vitro. The live vaccine has a good effect on the field of curing active tuberculosis or latent tuberculosis infection caused by proliferative mycobacterium tuberculosis, dormant mycobacterium tuberculosis and drug-resistant mycobacterium tuberculosis.

The invention relates to a pulmonary tuberculosis variation activity marker, a kit, a method and a model construction method, and the pulmonary tuberculosis variation activity marker is characterized in that the pulmonary tuberculosis variation activity marker is a plasma protein biomarker in a blood sample, and the plasma protein biomarker is a combined marker comprising one or more of C1QB protein and CCL19 (MIP3B) protein; a combined marker of C1QB and CCL19 (MIP3B) is used as a biomarker for detecting the activity of pulmonary tuberculosis for the first time, a rapid diagnosis model for detecting the activity of pulmonary tuberculosis is constructed, a new direction is provided for clinical diagnosis of tuberculosis, and as a technical conception for diagnosing the activity of pulmonary tuberculosis, the biomarker provides a new target spot for the diagnosis of the active tuberculosis; therefore, the invention overcomes the defects of low detection rate, long time consumption and the like of the existing active tuberculosis diagnosis, has the characteristics of good specificity and high sensitivity, and has good clinical application value for auxiliary diagnosis of tuberculosis activity.

The invention provides an active tuberculosis marker, a kit, a detection method and a model construction method. An active tuberculosis diagnostic marker is a plasma protein biomarker in a blood sample, wherein the plasma protein biomarker is a combined marker comprising one or more of APOA4 protein, CFH protein, CFHR5 protein, FGG protein and MBL2 protein. According to the invention, the combined marker of APOA4, CFH, CFHR5 and FGG derived from plasma protein is used as the biomarker for active tuberculosis detection for the first time, a rapid diagnosis model for active tuberculosis detection is constructed, and a new direction is provided for clinical diagnosis of tuberculosis. Therefore, the invention overcomes the defects of low detection rate, long time consumption and the like of the existing active tuberculosis diagnosis, has the characteristics of good specificity and high sensitivity, and has good clinical application value for the auxiliary diagnosis of the active tuberculosis.

The invention provides screening and application of active tuberculosis diagnosis molecules. In particular, the invention discloses high throughput screening of mycobacterium tuberculosis's important antigens and application thereof in active tuberculosis diagnosis. Based on a high throughput functional protein screening technology of GST fusion expression, 92 positive antigens recognizable by tuberculosis patients' serum are screened out, wherein 14 antigens present strongly positive reaction. Active tuberculosis serologic detection shows that the positive antigens have high sensitivity and specificity in detection of active tuberculosis. Specifically, TBGP1, TBGP2, TBGP3, TBGP4, TBGP5 and TBGP6 6 proteins form an antigen combination for tuberculosis serologic detection, and laboratory verification shows that the combination has high sensitivity and specificity in detection of mycobacterium tuberculosis, and has application value in tuberculosis diagnosis and monitoring.

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.