38 results about "Mycobacterium tuberculosis H37Rv" patented technology

The invention discloses a reagent for mycobacterium tuberculosis infection detection, clinical treatment effect tracking and antituberculous vaccine development. The novel mycobacterium tuberculosis derived reagent Rv3006 (LPPZ) prepared by a gamma-interferon gamma release assay and enzyme linked immunosorbent assay comprises antigen or polypeptide and analogues thereof, wherein terminal C to terminal N in the amino acid sequence of the Rv3006 antigen is as shown in SEQ ID NO:1. By utilizing the reagent, tuberculous patients in the active phase and tuberculous latent infectors can be well detected; the specific immune response of Rv3006 is reduced along with treatment development and disease condition remission, which indicates that the reagent can be used for well tracking the clinical treatment situation of a tuberculous patient, thus having a good clinical application prospect. Furthermore, the reagent has a protective effect on a mouse infected by a mycobacterium tuberculosis standard toxic strain H37Rv, thus having a good development prospect of antituberculous vaccines.

The invention discloses a kit of Mycobacterium tuberculosis and a detection method thereof. The kit contains an enzyme-linked immunosorbent assay (ELISA) plate, a biotin-labeled single-stranded DNA adaptor, horseradish peroxidase-labeled streptavidin and a chromogenic substrate. In the detection method provided by the invention, the biotin-labeled single-stranded DNA adaptor of Mycobacterium tuberculosis H37Rv is used as a probe and a method which is similar to the indirect enzyme-linked immunosorbent assay (ELISA) method is adopted to detect Mycobacterium tuberculosis; and Mycobacterium tuberculosis or sputum to be detected or a body fluid sample, and a control sample are added in the ELISA plate for incubation, Mycobacterium tuberculosis combined with the plate is trapped by the biotin-labeled single-stranded DNA adaptor and then washed, the horseradish peroxidase-labeled streptavidin is added, the substrate is added for developing, and the optical density (OD) value is detected by a microplate reader after developing when the absorption wavelengh is 450nm. When the kit provided by the invention detects Mycobacterium tuberculosis or the Mycobacterium tuberculosis or somatic antigen in sputum, blood and body fluid, the detection speed is high, the detection is convenient and safe, the detection time can be saved, the efficiency is high, the cost is low, the detection sensitivity and accuracy are high, and the application prospect is wide.

The invention belongs to the technical field of microbial genetic engineering, specifically relates to a construction method of a complete mycobacterium tuberculosis H37Rv genome ORF bacterial two-hybrid clone library and application thereof. The construction method is characterized by comprising the following steps: preparing a bacterial two-hybrid carrier pTRG-MCM delta suitable for constructing the complete mycobacterium tuberculosis H37Rv genome ORF clone library; obtaining all encoded genes of the mycobacterium tuberculosis H37Rv by a PCR reaction; accurately cloning all encoded genes of the mycobacterium tuberculosis H37Rv into the constructed bacterial two-hybrid carrier pTRG-MCM delta; and transforming Escherichia coli DH10B to obtain the complete mycobacterium tuberculosis H37Rv genome ORF bacterial two-hybrid clone library.

The invention discloses propylene tethered ciprofloxacin-isatin hybrids as well as a synthetic method and an application thereof. The prepared ciprofloxacin-isatin hybrid 3d have strong resistance toall tested Gram-positive bacteria and Gram-negative bacteria (including pathogens showing strong drug resistance clinically), and efficacy of 3d is equivalent to or even better than the parent ciprofloxacin or levofloxacin; antibacterial activity of hybrid 3b (MIC: 0.10 and 0.5 mu g / mL) for an mycobacterium tuberculosis H37Rv strain is 4 times and 8 times that of ciprofloxacin (MIC: 0.78 mu g / mL)and rifampicin (MIC: 0.39 mu g / mL), and antibacterial activity of the hybrid 3b for multi-drug resistant tuberculosis is 4->256 times that of three reference drugs including ciprofloxacin (MIC: 2.0 mug / mL), rifampicin (MIC: 32 mu g / mL) and isoniazid (>128 mu g / mL). The hybrids 3b and 3d with lower cytotoxicity (CC50: 64 and 256 mu g / mL) show acceptable metabolic stability and in-vivo PK (pharmacokinetics) characteristics.

The invention relates to a coding gene of Mycobacterium tuberculosis H37Rv. The coding gene can be used as a standard gene for molecular identification of a Mycobacterium tuberculosis complex group and can be used for molecular identification and clinical detection of the Mycobacterium tuberculosis complex group.

The invention belongs to the field of biomedicines, and relates to an application of vitamin C (ascorbic acid), as a single component, in preparation of medicines for treating and preventing tuberculosis. Through cytobiology and pharmacology experiments with combination of animal experiments, anti-tuberculosis pharmaceutical concentration of the vitamin C is determined; and on the basis of regulatory mechanism on oxidative stress status of mycobacterium tuberculosis after infection by macrophages, interference test is carried out, wherein a result proves that high-dose vitamin C can kill the mycobacterium tuberculosis, so that inhibition effect is achieved through NAC and catalase; the vitamin C treats the macrophages to generate H2O2, wherein functional effect is achieved with the signalchannel being same as H2O2; large-dose vitamin C kills mycobacteria, wherein inhibition function is achieved through a glutathione precursor NAC and CAT; high-dose vitamin C can infect the TNF-alpha signal pathway induced by RAW 264.7 cells via mediation of Bacillus Calmette-Guerin vaccine and H37Rv, thus achieving sterilization effect and influence on body immunity. The invention provides effective treatment strategy for clinical therapy and drug resistance of tuberculosis and tuberculosis in incubation period.

The invention relates to a specific electrochemical immunosensor for tuberculosis serodiagnosis, and belongs to the field of important epidemic prevention and cure. The immunosensor is a novel biosensor which is constructed through combining an immunoassay and a high sensitive sensing technology, and is applicable for analyzing and studying trace quantity of immunogenicity substance. According to the present invention, based on preparatory antigen marking and screening work, mycobacterium tuberculosis Rv2175c gene encoding protein (hereinafter referred to as specific antigen) is adopted as a specific antigen, a high sensitive electrochemical immunosensor is constructed, an antibody recognized by the specific antigen is detected in serum to realize the tuberculosis serodiagnosis. A method for the tuberculosis serodiagnosis comprises the following steps: (1) cloning the Rv2175c gene of the mycobacterium tuberculosis H37Rv; (2) heterologous expressing the purified specific antigen in escherichia coli; (3) preparing antibody serum on rabbits by using the specific antigen to be adopted as a positive control; (4) preparing the electrochemical immunosensor; (5) detecting the serum of healthy people and the serum of tuberculosis patients. With the present invention, the healthy people and the tuberculosis patients can be distinguished well through difference of current changing detected by the immunosensor, such that a purpose of serodiagnosis is achieved.

The invention discloses a novel small molecule compound and a preparation method and application thereof in preparation of mycobacterium drugs for resisting drug-resistant mycobacterium tuberculosis and like. The novel mycobacterium-resistant small molecule has a structural general formula (I), and the small molecule compound disclosed by the invention has a relatively good inhibition effect on mycobacteria. The antibacterial activity of mycobacterium smegmatis, mycobacterium tuberculosis H37Rv, H37Ra and drug-resistant mycobacterium tuberculosis are determined, and results show that the smallmolecule compound has good antibacterial activity on mycobacterium smegmatis, mycobacterium tuberculosis H37Rv, H37Ra and drug-resistant mycobacterium tuberculosis, and has an application prospect inpreparation of mycobacterium tuberculosis-resistant drugs.

The invention relates to a mycobacterium tuberculosis H37Rv coding gene which can serve as a standard gene for molecular identification of mycobacterium tuberculosis complex and is applied to the molecular identification and clinical detection of the mycobacterium tuberculosis complex.

The invention relates to a coding gene of Mycobacterium tuberculosis H37Rv. The coding gene can be used as a standard gene for molecular identification of a Mycobacterium tuberculosis complex group and can be used for molecular identification and clinical detection of the Mycobacterium tuberculosis complex group.

The invention relates to a coding gene of Mycobacterium tuberculosis H37Rv. The coding gene can be used as a standard gene for molecular identification of a Mycobacterium tuberculosis complex group and can be used for molecular identification and clinical detection of the Mycobacterium tuberculosis complex group.

The invention relates to a micro-molecule polypeptide. An amino acid sequence of a mature peptide of the micro-molecule polypeptide is as shown in SEQ ID No.1. The invention further discloses an application of the micro-molecule polypeptide to preparation of drugs for resisting mycobacterium tuberculosis. The micro-molecule polypeptide for resisting the mycobacterium tuberculosis stemmed from mycobacteriophage is small in molecular weight and low in synthetic cost and has obvious effects on resisting the mycobacterium tuberculosis H37Rv and the function of killing H37Rv resisting rifampin, other normal gram-positive bacteria, gram-negative bacteria and fungi are unaffected, and the micro-molecule polypeptide has a good application prospect in preparation of the drugs for resisting the mycobacterium tuberculosis.

The invention discloses a high throughput screening method of a mycobacterium tuberculosis alpha-D-glucose-1-thymine phosphate transferase inhibitor. A method for rapidly and accurately measuring the activity of the mycobacterium tuberculosis alpha-D-glucose-1-thymine phosphate transferase is established by the following steps of: cloning an rmlA gene from a mycobacterium tuberculosis H37Rv strain genome by using the molecular cloning technology; simultaneously creating an Escherichia coli engineering strain which highly expresses alpha-D-glucose-1-thymine phosphate transferase; and purifying the engineering strain to obtain mycobacterium tuberculosis alpha-D-glucose-1-thymine phosphate transferase by using affinity chromatography.. The transferase activity measuring method screens a molecular model of the mycobacterium tuberculosis alpha-D-glucose-1-thymine phosphate transferase inhibitor and is used for the high throughout screening of the mycobacterium tuberculosis alpha-D-glucose-1-thymine phosphate transferase inhibitor in combinational compound libraries, traditional Chinese medicine and natural products.

The invention relates to a coding gene of Mycobacterium tuberculosis H37Rv. The coding gene can be used as a standard gene for molecular identification of a Mycobacterium tuberculosis complex group and can be used for molecular identification and clinical detection of the Mycobacterium tuberculosis complex group.

The invention belongs to the technical field of medicines, and concretely relates to 3-aryl-6-formamidopyrazolo[1,5-a]pyrimidine compounds and an application thereof. The compounds are six 2,7-dimethyl-3-aryl-6-formamidopyrazolo[1,5-a]pyrimidine compounds prepared by preparing ethyl 2-dimethylaminomethylene-3-oxobutyrate (1) through a reaction of ethyl acetoacetate and N,N-dimethylformamide dimethyl acetal and carrying out cyclization, hydrolysis and amidation on the compound (1) and 3-methyl-4-(2,4-dichlorophenyl)-5-aminopyrazole, and the structure of the compounds is characterized by nuclear magnetic resonance hydrogen spectrum and infrared spectroscopy. A result of detection of the compounds by an MABA method shows that the synthesized compounds have an inhibition effect on Mycobacterium tuberculosis H37Rv. A pharmacological activity screening detection result shows that parts of the compounds have a very good inhibition effect on the Mycobacterium tuberculosis H37Rv and Mycobacterium phlei M. phlei1180, and has good anti-tuberculosis exploitation and application prospect.

The invention relates to building for a femoral epicondyle tuberculosis animal model, in particular to building of a rabbit femoral epicondyle tuberculosis animal model by using an H37Rv tuberculosisstrain. The model is an ideal platform for evaluating the effect of a drug delivery system in vivo. According to the method, the rabbit femoral epicondyle tuberculosis animal model is built by using the H37Rv tuberculosis strain, and the model is the ideal platform for evaluating the effect of the drug delivery system in vivo. The effectiveness and feasibility of the model are evaluated through gross specimen observation, Micro-CT examination, histological examination, mycobacterium culture and DNA micro-array testing. The femoral epicondyle animal model has the following advantages that the trauma is small in the preparation process, the operation is safe, and the model facilitates implantation of a material.

The invention relates to a mycobacterium tuberculosis H37Rv coding gene, which can be used as a standard gene for molecular identification of mycobacterium tuberculosis complex, and is used for molecular identification and clinical detection of mycobacterium tuberculosis complex.

The invention relates to a mycobacterium tuberculosis H37Rv new gene Rv2203c (-|2468297-2469040|) and a coding protein thereof. The protein coded by Rv2203c partially or completely has B cell antigenicity, and can be individually or jointly used for clinical rapid screening of tuberculosis patients and protection of human bodies from mycobacterium tuberculosis infection.

The invention relates to a novel coding gene Rv3350 (-|3760359-3761516|) of mycobacterium tuberculosis H37Rv and a coding protein of the novel coding gene Rv3350(-|3760359-3761516|). Part or all of protein encoded by Rv3350 has B cell antigenicity, and can be independently or jointly used for clinical rapid screening of pulmonary tuberculosis patients and prevent human bodies from being infected with the mycobacterium tuberculosis.

The invention discloses a mycobacterium tuberculosis fusion protein AR2, a construction, expression and purification method and application thereof. The construction, expression and purification method comprises the following steps: S1, extracting gene sequences coded by Rv1988 and Rv1886c from a whole genome sequence of mycobacterium tuberculosis H37Rv; S2, constructing a pET30b-Rv1988 recombinant plasmid and a pET28a-Rv1886c recombinant plasmid by taking pET28a and pET30b as carriers; S3, synthesizing Rv1988-Rv1886c, namely, a fusion protein AR2, according to the extracted gene sequences of the Rv1988 and the Rv1886c; S4, by taking the plasmid pET28a as a carrier, constructing a recombinant plasmid pET28a-AR2; S5, transferring the recombinant plasmids constructed in the step S2 and the step S4 into an expression vector E.coli BL21 strain; and S6, expressing and purifying the fusion protein AR2 and subcomponent proteins rRv1988 and rRv1886c. According to the invention, the fusion protein AR2 and an adjuvant are combined to construct a tuberculosis subunit vaccine, the vaccine can induce strong cellular immune response in a mouse body and promote generation of memory T cells, and the tuberculosis subunit vaccine is expected to become a more effective novel candidate vaccine for tuberculosis in the later period.

Relating to a vaccine preparation method, the invention provides a preparation method of a Mycobacterium tuberculosis Ag85A oral DNA vaccine. The method comprises: designing a pair of PCR primers according to a gene sequence of Mycobacterium tuberculosis Ag85A, taking the DNA of a human Mycobacterium tuberculosis H37Rv standard virulent strain as a template, conducting PCR amplification to obtain an Ag85A gene, subjecting a recycled PCR product to enzyme digestion by restriction endonuclease Xhol and BamHI, then connecting the product with a eukaryotic vector pCDNA3.1+ through a T4DNA ligase action, proving an obtained positive clone as the Ag85A gene by a DNA sequencing identification, cloning the Ag85A gene to the downstream of a CMV promoter in the vector pCDNA3.1+, constructing a recombinant plasmid pCDNA3.1+ / Ag85A, which is then transformed into Escherichia coli and amplified, thus obtaining the Mycobacterium tuberculosis Ag85A oral DNA vaccine. The method lays a foundation for clinical application research of oral DNA vaccines.

The invention relates to a coding gene of Mycobacterium tuberculosis H37Rv. The coding gene can be used as a standard gene for molecular identification of a Mycobacterium tuberculosis complex group and can be used for molecular identification and clinical detection of the Mycobacterium tuberculosis complex group.

The invention relates to an application of a 5- (3'-Indolyl)-oxazole compound as shown in formula I in preparing drugs for inhibiting mycobacterium tuberculosis and curing tuberculosis, and in the formula I, R represents a liner C2-C10 saturated alkane substituent group or a C2-C10 saturated alkane substituent group containing branched chains, and R is preferably methyl or ethyl. A microplate Alamar Blue method is used to test the antibacterial effect of the 5- (3'-Indolyl)-oxazole compound on mycobacterium tuberculosis H37Rv, which shows that the compound has mycobacterium tuberculosis resistance.