605 results about "Tuberculosis mycobacterium" patented technology

Disclosed are compositions and methods for isolating, detecting, amplifying, and quantitating Mycobacterium-specific nucleic acids in a sample. Also disclosed are compositions and diagnostic kits comprising Mycobacterium IS6110-specific oligonucleotide amplification primers and labeled oligonucleotide detection probes that specifically bind to the amplification products obtained therefrom. Also disclosed are compositions and methods for the isolation and characterization of nucleic acids that are specific to one or more tubercular pathogens, including Mycobacterium tuberculosis, in particular, from a wide variety of samples including those of biological, environmental, clinical and/or veterinary origin.

The invention provides methods and compositions for use in identifying inhibitors of biochemical pathways important for persistent infection, allowing the identification and/or design of improved therapeutics for treating persistent infections by pathogenic microbes. Particularly disclosed is the importance of the glyoxylate shunt to the persistent phase of various infectious agents, including Mycobacteria, such as M. tuberculosis, and the identification of preferred targets for drug development, including the enzymes isocitrate lyase (ICL) and malate synthase. Crystals and three-dimensional structures of M. tuberculosis ICL, without ligand and in complex with two inhibitors are also disclosed, for exemplary use in the design of inhibitors and therapeutic agents.

The invention relates to a double-marking probe detecting and melting curve analyzing method used for identifying mycobacterium strains and a kit using the method to identify various mycobacterium strains at the same time. The kit provided by the invention comprises a primer capable of designing mycobacterium 16S rRNA, a double-marking oligonucleotide probe capable of identifying mycobacterium strains, and a thermostable DNA polymerase without 5' nuclease activity. The detecting method and the kit, which are provided by the invention, can detect and/or identify 24 kinds of common mycobacteria. The method and the kit can judge the results according to melting peaks of different Tm values produced by the sequence hybridization of the probe and the different strains, meanwhile rapidly and accurately detect the mycobacteria, and identify the mycobacteria, non-mycobacteria, mycobacterium tuberculosis compounding groups and nontuberculosis mycobacteria, and the result of identifying the mycobacteria can be reported after 3-4 hours, thus the method and the kid can assist the clinical diagnosis, and guide efficient clinical chemotherapy at the early stage.

The present invention is based on the identification and characterization of a number of M. tuberculosis derived novel proteins and protein fragments (SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 17-23, 42, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72-86, 88, 90, 92, 94, 141, 143, 145, 147, 149, 151, 153, and 168-171). 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. Another part of the invention is based on the surprising discovery that fusions between ESAT-6 and MPT59 are superior immunogens compared to each of the unfused proteins, respectively.

The present invention relates to novel substituted quinoline derivatives according to the general Formula (Ia) or the general Formula (Ib)

the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the tautomeric forms thereof and the N-oxide forms thereof. The claimed compounds are useful for the treatment of mycobacterial diseases, particularly those diseases caused by pathogenic mycobacteria such as Mycobacterium tuberculosis, M. bovis, M. avium and M. marinum. In particular, compounds are claimed in which, independently from each other, R¹ is bromo, p=1, R² is alkyloxy, R³ is optionally substituted naphthyl or phenyl, q=1, R⁴ and R⁵ each independently are hydrogen, methyl or ethyl, R⁶ is hydrogen, r is equal to 0 or 1 and R⁷ is hydrogen. Also claimed is a composition comprising a pharmaceutically acceptable carrier and, as active ingredient, a therapeutically effective amount of the claimed compounds, the use of the claimed compounds or compositions for the manufacture of a medicament for the treatment of mycobacterial diseases and a process for preparing the claimed compounds.

The present invention discloses a screening method of a compound for inhibiting mycobacterium tuberculosis. The method comprises the following steps: (1) establishing a screening model adopting tyrosine phosphatase as a target point: adopting genome DNA of tubercle bacillus H37 Rv as a template, adopting a PCR technology to clone tyrosine phosphatase gene, transforming host cells, culturing the transformant to obtain recombinant tyrosine phosphatase from the culture, carrying out enzyme activity analysis, and establishing the screening model adopting tyrosine phosphatase as the target point; and (2) adopting the screening model established in the step (1) to screen a compound providing inhibition activity for the tyrosine phosphatase. With the method of the present invention, the tyrosine phosphatase inhibitor with anti-mycobacterium tuberculosis effect can be rapidly and efficiently screened. In addition, the present invention further discloses a compound screened by using the screening method, and uses of the compound in preparations of drugs for treatments of diseases, wherein the inducement or one of the inducements of the diseases is the tyrosine phosphatase.

The invention discloses streptomyces atratus and application of cyclic peptide compounds with the same to preparing mycobacterium tuberculosis resistant medicines. A structural formula of the cyclic peptide compounds is shown. A preservation number of the streptomyces atratus SCSIO Zh16 is CGMCC No.12198. The streptomyces atratus and the application have the advantages that the six cyclic peptide compounds are obtained from fermentation cultivation substances of the streptomyces atratus SCSIO Zh16 by means of separation, the cyclic peptide compound 6 is high in mycobacterium tuberculosis resistant activity, obvious effects of inhibiting mycobacterium tuberculosis can be realized by the cyclic peptide compound, accordingly, the cyclic peptide compounds can be used for preparing anti-tuberculosis medicines and can be used for treating tuberculosis, alternative compounds can be provided for developing novel anti-tuberculosis medicines, and the streptomyces atratus and the application have important significance on developing marine medicinal materials in China.

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 polypeptide.

The invention discloses a reagent for detecting tubercle bacillus infection in vitro and a method thereof. The reagent comprises M233 polypeptide represented by SEQ ID No.1; and cytokine released from T cells is detected by contacting the M233 polypeptide or an analog thereof with the T cells of a tubercle bacillus host to determine whether the T cells identify the M233 polypeptide or the analog thereof. The reagent has the advantages of high sensitivity, good specificity, being free from interference of BCG vaccine and non-tuberculosis mycobacteria vaccine, being capable of detecting active pulmonary tuberculosis patients, the patients with dormant infection and healthy persons who contact with mycobacterium nontuberculosis. The reagent and the method are especially applicable to detecting tuberculosis and/or dormant infection thereof for Chinese people.

The invention relates to a method for detecting multi-drug resistance of Mycobacterium tuberculosis, which aims at detecting the resistance of the Mycobacterium tuberculosis to isoniazid and rifampicin at the same time and has the characteristics of high specificity and sensitivity, quickness in detection, and easiness and convenience in operation. The technical scheme is as follows: the method comprises the following steps: A, establishing PCR (polymerase chain reaction) templates of an MTB clinical strain and a standard strain H37Rv; B, designing three pairs of primers of katG, inhA and rpoB gene fragments which are closely related to the isoniazid resistance and the rifampin resistance, performing mPCR amplification on katG, inhA and rpoB genes of the clinical strain, and performing PCR amplification on katG, inhA and rpoB genes of the clinical strain and the standard strain H37Rv; and C, detecting the mutation conditions of related INH-resistant and RFP-resistant 3 genes of the clinical strain at the same time by a single-strand conformation polymorphism detection method.

The current invention provides a series of bicyclic nitroimidazole- substituted phenyl oxazolidinones in which a bicyclic nitroimidazole pharmacophore is covalently bonded to a phenyl oxazolidinone, their pharmaceutical compositions, and the method of use of the compositions for prevention and treatment of bacterial infections. The bicyclic nitroimidazole-substituted phenyl oxazolidinones possess surprising antibacterial activity against wild- type and resistant strains of pathogens, and are therefore useful for the prevention, control and treatment of a number of human and veterinary bacterial infections caused by these pathogens, such as Mycobacterium tuberculosis.

The invention relates to a tuberculosis antibody multiple antigen ELISA detection kit and a preparation method thereof, which pertains to the field of tuberculosis medical immunology diagnostic techniques and mainly uses detection antigen, enzyme-linked antihuman IgG antibodies, substrates, positive control serum of tuberculosis patients, control serum of normal person, calf serum and polystyrene microplates to form the kit, wherein, the detection antigen adopts the mycobacterium tuberculosis complex strains of lipid Arabian mannose (LAM), 38kD and 16kD to be combined with arbitrary one or more than one mycobacterium tuberculosis recombinant proteins in the recombinant proteins of MPT63, MTB48 and CFP10-ESAT6. The mycobacterium tuberculosis has high sensitivity, strong specificity and complementarity, can be used for detecting specific antitubercular antibodies in such body fluid samples as serum, hydrothorax and the like, and assisting the diagnosis and differential diagnosis of tuberculosis.

The invention discloses a mycobacterium tuberculosis TB10.4-F1 fusion protein vaccine and a preparation method thereof. The preparation method comprises the following steps of: connecting the mycobacterium tuberculosis TB10.4 protein gene with the F1 protein gene of respiratory syncytial virus protein, transferring into a colibacillus expressing strain, inducing by IPTG (isopropyl beta-D-1-thiogalactopyranoside) to express the TB10.4-F1 fusion protein, and acquiring the protein with higher purity by metal ion affinity chromatography. The fusion protein has stronger immunogenicity, can excite more balanced Th1/Th2 immune response, and can prevent infections of the mycobacterium tuberculosis and the respiratory syncytial virus.

The invention relates to a reagent kit for diagnosing enzyme linked immune spots infected by tubercle bacillus and a method for preparing a specific antigen. The reagent kit comprises a reagent kit body and a detection reagent arranged in the kit body; the detection reagent comprises a positive standard solution, a chromogenic agent, a concentrated detergent and a diluent; the detection reagent also comprises a specific antigen of mycobacterium tuberculosis, a coated INF-gamma resistant antibody and an enzyme label secondary antibody of a vector protein conjugate; the specific antigen of mycobacterium tuberculosis is fusion protein expressed by an ESAT-6 gene and an EIS gene of the mycobacterium tuberculosis; and the INF-gamma resistant antibody is a murine IgG antibody. The method for preparing the specific antigen comprises the following steps: proper tubercle bacillus special antigen ESAT-6 and EIS are combined; and the prepared recombining fusion protein contains main antigenic determinants of two antigens. The recombining fusion protein definitely contains a T cell epitope suitable for the limitation of different HLA, and does not influence results by different groups and different HLA distribution. Clinical tests on tuberculosis patients and healthy people of different groups show that the reagent kit has more excellent specificity and sensitivity than a current similar product.

The invention discloses a detection method of mycobacterium tuberculosis pyrazinamide drug resistance, comprising the following steps: A. pyrolyzing a template of mycobacterium tuberculosis used for polymerase chain reaction (PCR); B. designing a PCR primer, and augmenting a full-length pyrazinamide (pncA) gene; C. concentrating the augmented pncA gene segment and quantifying; D. expressing pncA enzyme by adopting a wheat germ acellular expression system; and F. determining the expressed pncA enzymatic conversion pyrazinamide into the activity of pyrazine acid; comparing the pncA enzymatic activities of a sensitive strain and a drug resistance strain of the expressed tubercle bacillus standard; and determining the drug resistance of the mycobacterium tuberculosis. The method is characterized in that the drug resistance caused by pncA gene mutation can be determined by a pair of primers, and simultaneously discloses a primer sequence for augmenting the full-length pncA gene from the tubercle bacillus gene group and being suitable for a wheat germ acellular expression system to express the mycobacterium tuberculosis pyrazinamide. By the detection method, the mycobacterium tuberculosis pyrazinamide drug resistance can be rapidly and sensitively detected in no need of bacterial culture.

The invention discloses a method and a kit for detecting drug resistance of mycobacterium tuberculosis. The invention provides special-purpose primers and special-purpose probes for identification of drug resistance of mycobacterium tuberculosis in a sample needing to be detected, wherein the special-purpose primers comprise multiple primer groups; each one of the primer groups comprises a commonprimer and multiple specific primers for amplification of alleles corresponding to all mutational sites of same-purpose genes; the special-purpose probes comprise multiple types of probes; and each one of the probes specifically binds with target sequences corresponding to the common primer and the specific primers of the primer group. Experiments of the invention prove that the special-purpose primers and the special-purpose probes can be combined by an efficient and sensitive duplex Taqman real-time fluorescent polymerase chain reaction (PCR) technology and a sequence specific primer (SSP) amplification technology so that drug resistance of mycobacterium tuberculosis in a detected sample can be detected in a short time.

The invention provides polypeptides encoded by open reading frames present in the genome of Mycobacterium tuberculosis but absent from the genome of BCG and diagnostic and prophylactic methodologies using these polypeptides.

Vaccination with the combination of Ag85B-TB10.4 and IC31® adjuvant generated a high amount of polyfunctional CD4⁺T cells expressing high levels of IFN-γ, TNF-α, and IL-2. This in turn led to significant protection against infection with M. tuberculosis in the mouse aerosol challenge model of tuberculosis. Both the immunogenicity of the vaccine and its ability to protect against TB infection was highly dependent on the antigen dose. Thus, whereas the standard antigen dose of 5 μg, as well as 15 μg, did not induce significant protection against M. tuberculosis, reducing the dose to 0.5 μg increased both the immunogenicity of the vaccine as well as its protective efficacy to a level comparable to that observed in BCG vaccinated mice. Thus, the IC31® adjuvant, with the specified antigen dose, can induce a strong protective Th1 response against M. tuberculosis.

Relating to the detection technology of resistant mutation of mycobacterium tuberculosis, the invention aims to provide a rapid, sensitive and specific method for detecting the resistant mutation of mycobacterium tuberculosis to rifampin and a kit thereof. The method comprises the steps of: extracting the DNA of a mycobacterium tuberculosis sample; according to the rpoB gene sequence (GenBank: L27989) where an RRDR (rifampin resistance determining region) of mycobacterium tuberculosis is located, designing primers and probes by means of the primer design software Primer Premier 5; constructing a double-tubed and double-colored real-time PCR (polymerase chain reaction) system, detecting the mutation of the RRDR of mycobacterium tuberculosis, comparing the melting point (Tm value) differences of melting curves between the detected sample and a positive control, and determining whether the sample mutates. With the primers designed at two ends of the RRDR of the rpoB gene, the method of the invention is of high specificity.

The invention relates to a joint detection method for the drug resistance of mycobacterium tuberculosis and the pyrazinamide thereof in a clinical sample, comprising the following steps of: A, extracting DNA (deoxyribonucleic acid) in the sample; B, detecting the mycobacterium tuberculosis via fluorescent quantitative PCR (polymerase chain reaction) amplification for a tuberculosis genome segment containing total-length pyrazinamide enzyme genes (pncA); and C, detecting the drug resistance of pyrazinamide for the mycobacterium tuberculosis positive sample, wherein the steps for detecting the drug resistance of the pyrazinamide are as follows: 1, performing PCR reaction once by taking the fluorescent quantitative PCR product as a template, so as to add an in-vitro expression element; 2, adding the PCR product in a cell-free expression system, so as to express a pyrazinamide enzyme; and 3, comparing the pyrazinamide enzyme activity of the sample with a standard strain, so as to give the drug resistance result of pyrazinamide. According to the invention, joint detection for the drug resistance of mycobacterium tuberculosis and the pyrazinamide thereof in the sample is realized by combining fluorescent quantitative PCR amplification for pncA genes with the enzyme activity of the in-vitro expressed pyrazinamide enzyme, without the need of bacterial culture and a DNA sequencer.