434 results about "Tuberculosis mycobacterium" patented technology

The invention provides an intracellular mycobacterium tuberculosis in-situ nucleic acid detection method based on CRISPR / Cas and application, and the method comprises the following steps: S1, designing crRNA and a primer pair of a mycobacterium tuberculosis target gene sequence, and synthesizing a single-stranded DNA fluorescent probe; s2, preparing a standard solution containing a target gene sequence, and incubating with a CRISPR / Cas12a system; s3, extracting nucleic acids of different types of bacteria, uniformly mixing the nucleic acids of different types of bacteria with the CRISPR / Cas12a system, and incubating; s4, incubating an attenuated strain of mycobacterium tuberculosis and the DNA fluorescent probe, infecting macrophages, and incubating to obtain infected cells; s5, electrically transferring a CRISPR / Cas12a system into the infected cells in S4 by utilizing an electroporation technology, incubating, and detecting the intensity of a fluorescence signal; and S6, performing quantitative and positioning analysis on the detected fluorescence signal to complete detection, and directly introducing a CRISPR / Cas12a system into the macrophage by adopting an electroporation technology to realize intracellular in-situ detection, thereby avoiding the operation complexity of a nucleic acid extraction link and the loss of detection sensitivity in the prior art.

The invention discloses an application of cryptotanshinone in preparation of an anti-mycobacterium tuberculosis drug, and relates to the technical field of bioengineering. The molecular formula of the valerianin disclosed by the invention is C19H20O3. The mycobacterium tuberculosis is a mycobacterium tuberculosis H37Rv strain. Cryptotanshinone is disclosed as a natural-source compound for the first time, has the dual advantages of high efficiency and low toxicity, has an obvious effect of resisting mycobacterium tuberculosis, provides a brand new candidate drug scheme with development potential for solving the difficulty of tuberculosis treatment, and has great scientific value and application prospect.

The invention discloses a method for rapidly detecting mycobacteria. The method comprises the following steps: using a primer pair for detecting the Mycobacterium tuberculosis, wherein the primer pair comprises an upstream primer and a downstream primer; the upstream primer comprises a nucleotide sequence as shown in SEQ ID NO: 1 or 2, and the downstream primer comprises a nucleotide sequence as shown in any one of SEQ ID NO: 3-6. According to the present invention, the RNA of the MTB viable bacteria in the biological sample can be rapidly and accurately detected with high sensitivity, the detection limit is reduced by 1000 times compared with the existing NASBA kit, the template requirement is reduced, and the better choice is provided for the instant detection of MTB.

The invention discloses a primer composition and application thereof. According to the invention, a group of primer compositions is obtained aiming at mycobacterium tuberculosis complex group identification and drug resistance related genes thereof. The primer composition can be combined with nucleic acid matrix-assisted laser desorption ionization time-of-flight mass spectrometry to detect a mycobacterium tuberculosis complex group, and can also detect the mutation conditions of 18 drug-resistant genes and 92 gene mutations related to drug resistance of 21 common first-line and second-line antituberculous drugs / types in the mycobacterium tuberculosis complex group. The medication is guided. Based on a MassARRAY technical platform, the primer composition has the advantages of good accuracy, high sensitivity, short required time and high efficiency when being used for detecting the mycobacterium tuberculosis complex group and the drug resistance thereof, is suitable for various sample types, can meet clinical detection requirements, is beneficial to clinical diagnosis and medication guidance of mycobacterium tuberculosis, and realizes precise treatment of pulmonary tuberculosis.

The present invention relates to a recombinant Mycobacterium bovis BCG strain comprising a plasmid having a short guide RNA (sgRNA) target sequence for knocking out the Mb3739 gene. Also provided are vaccine compositions for eliciting an immune response against Mycobacterium tuberculosis comprising a recombinant Mycobacterium bovis BCG strain engineered to activate the NOD-1 pathway. The recombinant Mycobacterium bovis BCG strain or vaccine composition can be used in a method against an immune response initiated by Mycobacterium tuberculosis. The invention also relates to a method for obtaining the recombinant bovine mycobacterium BCG strain.

Provided herein are nucleic acid vaccine constructs comprising synthetic polynucleotides encoding a Mycobacterium tuberculosis (Mtb) RelA-SpoT homolog (RSH) protein, RelMtb, or a functional portion, fragment, or variant thereof, conjugated to a macrophage inflammatory protein-3 alpha (MIP-3α) or other chemokine that binds to a chemokine receptor 6 (CCR6), or a functional portion, fragment, or variant thereof, or to an antibody, or antigen binding portion thereof, that binds to a CCR6. Methods for making the vaccine constructs and their use in prophylaxis and treatment of Mtb infections are also provided.

The invention discloses a detection kit and method for Mycobacterium tuberculosis complex and rifampicin drug-resistant mutation, and belongs to the technical field of biologication.A Taqman probe and a molecular beacon are combined, a fluorescent signal is generated based on a Taq enzyme hydrolysis fluorescent probe, the fluorescent signal is accumulated, and the molecular beacon is used for detecting the drug-resistant mutation of the Mycobacterium tuberculosis complex and the rifampicin drug-resistant mutation of the Mycobacterium tuberculosis complex and the rifampicin drug-resistant mutation. According to two technical principles that a molecular beacon and a fluorescent product are hybridized to generate a fluorescent signal, fluorescent PCR amplification is adopted for detection of one target spot in a monochromatic fluorescent channel, and a melting curve mode of a probe and a target gene is adopted for detection of the other target spot. Therefore, two target spots can be detected and analyzed in a monochromatic fluorescence channel at the same time. Single-tube multiple detection is achieved, and mycobacterium tuberculosis complex group detection and mycobacterium tuberculosis rifampicin drug-resistant mutation site detection can be achieved at the same time.

The invention belongs to the technical field of biology, and particularly relates to a primer probe composition and a kit for detecting lower respiratory tract infection pathogens. The primer probe composition is used for detecting mycobacterium tuberculosis, mycobacterium avium, mycobacterium abscessus, mycobacterium kansasii, pseudomonas aeruginosa, klebsiella pneumoniae, acinetobacter baumannii and haemophilus influenzae, and the primer probe composition is used for detecting mycobacterium tuberculosis, mycobacterium avium, mycobacterium abscessus, mycobacterium kansasii, pseudomonas aeruginosa, klebsiella pneumoniae, acinetobacter baumannii and haemophilus influenzae. The composition is one or more of a composition for detecting staphylococcus aureus, a composition for detecting streptococcus pneumoniae, a composition for detecting escherichia coli, a composition for detecting moraxella catarrhalis, a composition for detecting serratia marcescens, a composition for detecting mycoplasma pneumoniae, a composition for detecting candida albicans or a composition for detecting adenovirus; the RAA-RDB technology has the core advantages of rapidness, high sensitivity, low cost and easy operation, and is especially suitable for multiple pathogen screening and basic medical application.

The invention discloses identification and application of a mycobacterium tuberculosis bacteriophage susceptibility gene, and a gene target which affects mycobacterium bacteriophage infection efficiency is obtained through combined application of a mycobacterium tuberculosis CRISPR-KO library and a CRISPRi library and bacteriophage D29 infection screening. Reagents or drugs targeting phage susceptibility genes or encoded proteins thereof are screened based on the gene targets, and the reagents or drugs can knock out the phage susceptibility genes or inhibit expression of the phage susceptibility genes, or inhibit activity of the encoded proteins of the phage susceptibility genes, serve as phage synergists, or are combined with antibiotics to be used with phages, and can be used for preparing drugs for treating the phage susceptibility genes or the encoded proteins of the phage susceptibility genes. The bacteriophage-antibiotic composition has the advantages that a synergistic sterilization effect is achieved, a new strategy is provided for bacteriophage-antibiotic combined treatment of drug-resistant mycobacterium infection, rapid sterilization is expected to be achieved, the treatment course is shortened, and finally the target of rapid treatment of tuberculosis and other pathogenic mycobacterium infection is achieved.

The present invention provides a microfluidic detection chip for tuberculosis-specific cellular immunity, belonging to the technical field of Mycobacterium tuberculosis detection. The chip is primarily composed of a three-layer structure, assembled sequentially from bottom to top: a chip bottom sealing plate, a chip substrate, and a chip top sealing plate. The chip is equipped with detection units for multiple samples, capable of simultaneously performing IGRA detection of negative controls, positive controls, and a sample control of 1-8 test samples. Furthermore, the present invention provides an assembly process for a centrifugal microfluidic detection chip for tuberculosis-specific cellular immunity, as well as a detection method and application of chip-based IGRA using the chip. The chip-based IGRA demonstrated a 100% overall concordance rate between test results and those of conventional IGRA. Furthermore, the chip-based IGRA enables detection of very small numbers of test samples, while improving the throughput and speed of IGRA detection, thus showing significant application prospects for IGRA detection of large numbers of clinical samples.

The invention provides a compound as shown in a formula (I), a stereoisomer thereof, an optical isomer thereof, a pharmaceutically acceptable salt thereof, a crystal form thereof, an isotope derivative thereof, a prodrug thereof and a metabolite thereof. The solvate or hydrate of the compound can be used for preparing a pharmaceutical composition for treating and / or preventing diseases related to mycobacterium tuberculosis and / or nontuberculous mycobacterium. The compound provided by the invention has an excellent bactericidal effect and strong specificity, has no cross resistance with the existing antituberculosis drugs, and provides a breakthrough direction for treatment of drug-resistant tuberculosis.

The invention discloses a combination of blocking agents for detecting drug resistance of mycobacterium tuberculosis. The combination comprises a first peptide nucleic acid blocking agent, a second peptide nucleic acid blocking agent, a third peptide nucleic acid blocking agent and a fourth peptide nucleic acid blocking agent. The first peptide nucleic acid blocker covers codons 511 and 513 of the rpoB gene. The second peptide nucleic acid blocker covers the codon 516 of the rpoB gene. And the third peptide nucleic acid blocking agent covers the codon 526 of the rpoB gene. And the fourth peptide nucleic acid blocking agent covers codons 531 and 533 of the rpoB gene. The respective sequences of the first peptide nucleic acid blocker, the second peptide nucleic acid blocker, the third peptide nucleic acid blocker, and the fourth peptide nucleic acid blocker match the wild type of the corresponding sequence of the rpoB gene. The invention also provides a kit and a method for detecting the drug resistance of mycobacterium tuberculosis.

The invention discloses application of Rv2610 in preparation of a medicine for resisting mycobacterium tuberculosis infection, and belongs to the technical field of biology. The invention finds that the Rv2610 can promote the adhesion capacity of mycobacterium tuberculosis to macrophages and inhibit the expression of early inflammatory molecules TNF-alpha and IL-1beta of the macrophages, and in a mouse infection model, the Rv2610 promotes the colonization of mycobacterium in the lung in vivo. The Rv2610 can be used as a new drug target for resisting tuberculosis infection, and an inhibitor of the Rv2610 can be used for preparing a drug for resisting mycobacterium tuberculosis infection. The invention provides new tools and ideas for clinical prevention and treatment of tuberculosis, particularly has wide prospects in the aspects of development, clinical application and the like of antituberculosis drugs and vaccines, and can also be applied to the field of scientific research or guide development of inhibitors for inhibiting mycobacterium tuberculosis from infecting host macrophages. The invention has important application value for obtaining new anti-tuberculosis drug targets and screening new drugs.

The invention relates to the technical field of biological medicines, in particular to a tuberculosis vaccine antigen and a vaccine. In particular to application of a mycobacterium tuberculosis Rv1787 gene in preparation of a vaccine for preventing and / or treating tuberculosis, in the application, the sequence of the Rv1787 gene or an encoding product thereof is defined, and the Rv1787 gene and the encoding product thereof serve as antigens, have mycobacterium tuberculosis specificity, and can be used for preparing a vaccine for preventing and / or treating tuberculosis. According to the mycobacterium tuberculosis vaccine, identification and response of an organism immune system to mycobacterium tuberculosis can be accurately stimulated, non-specific immune response to other normal tissues is avoided, a brand new and specific molecular target spot is provided for preventing and treating tuberculosis, and the limitation that a traditional vaccine depends on a single antigen or an attenuated strain is broken through. A specific expression product of the gene can stimulate directional recognition and attack of an organism immune system on mycobacterium tuberculosis, the pertinence and effectiveness of vaccines are remarkably improved, and a new path is provided for solving the problem of tuberculosis prevention and treatment.

The present invention provides an electrochemical aptamer sensor for detecting Mycobacterium tuberculosis ESAT‑6 antigen. The sandwich electrochemical biosensor uses a new type of MXene@Hemin‑AuNPs nanozyme as a signal amplifier and N‑CNTs‑AuNPs as a sensing interface to detect the Mycobacterium tuberculosis-specific antigen ESAT‑6 in the serum of tuberculosis patients, providing a new method for the diagnosis of tuberculosis.

This invention provides a data storage method and system for tuberculosis detection, relating to the field of tuberculosis detection technology. The data storage method for tuberculosis detection includes the following steps: S1. Collecting whole-genome sequencing data of Mycobacterium tuberculosis, host serum IgG titer, and drug sensitivity test results; S2. Calculating genetic distance D based on a reverse evolution model to generate four-dimensional spatiotemporal coordinates (t, x, y); S3. Performing data partitioning and storage based on the drug target barrier value β; S4. Generating dynamic metadata using a host-pathogen dynamics model and compressing and storing it. This invention implements a dynamic storage entropy adjustment algorithm at the hardware and software collaborative level, continuously optimizing the matching efficiency of data compression and physical storage. This results in an intelligent data hub that can perceive the evolutionary pulse of pathogens and autonomously optimize resources, providing support for clinical tuberculosis prevention and control decisions with temporal depth, spatial correlation, and risk evolution.

Provided here are nucleic acid constructs containing a Mycobacterium tuberculosis genome comprising a mutation (such as a deletion) of the sigH gene (AsigH) and a mutation (such as a deletion) of at least one additional gene, and mutant Mycobacterium tuberculosis encoded by the nucleic acid constructs. Also provided are methods of making such nucleic acid constructs and Mycobacterium tuberculosis mutants, and uses thereof. The construct can stimulate an immune response against Mycobacterium tuberculosis in a subject, and can be used as live attenuated vaccines.

Disclosed are an mRNA pharmaceutical composition for preventing and treating tuberculosis and use thereof. The mRNA pharmaceutical composition comprises: an mRNA molecule encoding a Mycobacterium tuberculosis antigen, and a pharmaceutically acceptable excipient. The Mycobacterium tuberculosis antigen comprises the following antigen components: at least one early-secreted antigen of Mycobacterium tuberculosis or an immunologically active fragment thereof; PE / PPE family antigen WAG22 of Mycobacterium tuberculosis or an immunologically active fragment thereof; and at least one latent-related antigen of Mycobacterium tuberculosis or an immunologically active fragment thereof. The mRNA pharmaceutical composition does not comprise or further comprises an mRNA molecule encoding a cytokine. The pharmaceutical composition is used for preparing a tuberculosis vaccine, which may serve as a prophylactic vaccine for preventing latent activation or as a therapeutic drug for treating active tuberculosis, exhibiting a significant inhibitory effect on Mycobacterium tuberculosis.

The invention discloses a CRISPR-Cas13a (clustered regularly interspaced short palindromic repeats-CRISPR-Cas13a) double crRNA (ribonucleic acid) system which is established by using ERASE test paper and is used for detecting the rifampicin resistance of mycobacterium tuberculosis on site. The CRISPR-Cas13a double crRNA-system comprises a Cas13a protein and crRNA, or a complex formed by the Cas13a protein and the crRNA; the crRNA comprises an anchoring sequence and a guide sequence; the anchoring sequence is used for being combined with Cas13a protein; the guide sequence is used for targeting mutation target sequences of sites 531, 526, 516 and 513 in a mycobacterium tuberculosis drug-resistant gene rpoB gene; the target sequence of the mycobacterium tuberculosis is located at the 2317th site to the 2597th site of the rifampicin drug-resistant gene rpoB. On the basis of a CRSPR-Cas13a nucleic acid detection technology, result interpretation is performed through RAA amplification in combination with a line elimination method (ERASE) lateral flow chromatography test paper, and experiments prove that the kit can target high-frequency mutation sites of four rifampicin drug-resistant genotypes at the same time, has good detection sensitivity and specificity, and can be used for detecting the rifampicin drug-resistant genotypes of the rifampicin drug-resistant genotypes of the rifampicin drug-resistant genotypes of the rifampicin drug-resistant genotypes. The method can realize rapid, efficient, simple and convenient detection of mycobacterium tuberculosis rifampicin drug-resistant nucleic acid, has a sensitivity of 10 copies / [mu] L, and has an important application value.

The invention discloses a primer composition for detecting drug resistance of mycobacterium tuberculosis, a related product and application, and belongs to the field of pathogen detection. The primer composition comprises a primer pair aiming at an rpoB gene, a primer pair aiming at a katG gene, a primer pair aiming at an inhA gene, a primer pair aiming at an ahpC gene, a primer pair aiming at a gyrA gene and a primer pair aiming at a gyrB gene. According to the invention, mutation of mycobacterium tuberculosis complex and rifampicin, isoniazide and quinolone drug resistance related genes rpoB, katG, inhA, ahpC, gyrA and gyrB genes in sputum of a tuberculosis patient can be detected at one time, single-tube multiple high-sensitivity detection is realized, wild genes and mutant genes are accurately distinguished, and serious cross reaction is avoided.

The present invention relates to novel rufamycin analogs that target the ClpC1 protease. The compounds of the present invention have antibacterial activity, e.g., with anti-tuberculosis properties, and are useful in the treatment of e.g. Tuberculosis.

The invention provides an mRNA (messenger ribonucleic acid) pharmaceutical composition for preventing and treating tuberculosis and application of the mRNA pharmaceutical composition. The mRNA pharmaceutical composition for preventing and treating tuberculosis comprises mRNA molecules for coding mycobacterium tuberculosis antigens and pharmaceutically acceptable auxiliary materials, the mycobacterium tuberculosis antigen comprises the following antigen components: at least one mycobacterium tuberculosis early secretion antigen or an immunocompetence fragment thereof; a mycobacterium tuberculosis PE / PPE family antigen Rv3872 or an immunocompetence fragment thereof; and at least one mycobacterium tuberculosis latent associated antigen or an immunocompetent fragment thereof; the mRNA pharmaceutical composition does not include or further includes an mRNA molecule encoding a cytokine. The pharmaceutical composition provided by the invention is used for preparing tuberculosis vaccines, can be used as a prophylactic vaccine for preventing latent activation, can also be used as a therapeutic drug for treating active tuberculosis, and has a remarkable inhibition effect on mycobacterium tuberculosis.

The invention is applicable to the field of genetic engineering, and provides a mycobacterium tuberculosis Mce1A-LS protein nanoparticle, a preparation method and application thereof, the mycobacterium tuberculosis Mce1A-LS protein nanoparticle is formed by sequence fusion of mycobacterium tuberculosis Mce1A protein and LS protein, and the amino acid sequence of the mycobacterium tuberculosis Mce1A-LS protein nanoparticle is shown as SEQ ID NO: 2 in a sequence table. The Mce1A-LS protein nanoparticle provided by the invention is high in immunogenicity, the protein Mce1A with high immunogenicity can induce to generate a synergistic immune effect and is high in safety, and the expressed fusion protein is non-toxic and harmless, so that the biosafety is high, in addition, the immune effect of the Mce1A-LS protein nanoparticle is good, and after the Mce1A-LS protein nanoparticle is used for immunizing a mouse, the immunogenicity of the Mce1A-LS protein nanoparticle is greatly improved. Strong body fluid and / or cellular immune response can be generated, which indicates that after immunization, a specific immune effect can be generated in a mouse body.

Provided is a method for rapidly detecting Mycobacterium tuberculosis. The method comprises using a primer pair for detecting Mycobacterium tuberculosis, which primer pair comprising an upstream primer and a downstream primer; wherein the upstream primer comprises a nucleotide sequence as shown in SEQ ID NO: 1 or 2, and the downstream primer comprises a nucleotide sequence as shown in any one of SEQ ID NOs: 3-6. The method can rapidly and accurately detect the RNA of MTB viable bacteria in a biological sample with high sensitivity, reduces the detection limit by up to 1000 times compared with existing NASBA kits, reduces the requirements for templates, and provides a better option for the real-time detection of MTB.

The application belongs to the technical field of biological medicine, and particularly relates to a blood Mycobacterium tuberculosis complex free nucleic acid detection kit and a detection method. The application first proposes a kit, wherein a PCR detection reagent group and a CRISPR / Cas12a detection reagent group are arranged in the kit; the PCR detection reagent group comprises a forward primer and a reverse primer which target a complex target; the sequence of the forward primer is overlapped with or complementary to the recognition region of the crRNA sequence, or the sequence of the reverse primer is overlapped with or complementary to the recognition region of the crRNA sequence.

The invention discloses a detection system and method for identifying mycobacteria and drug resistance genes of the mycobacteria based on nanopore sequencing, and relates to the field of biological medicine, the detection system comprises a specific targeted enrichment module, a nanopore sequencing module and a biological information analysis module; the specific targeted enrichment module comprises a probe combination, and the probe combination covers a mycobacterium tuberculosis complex conservative identification gene, species-specific genes of common nontuberculous mycobacteria and mycobacterium leprosy, and full-length or partial sequences of drug resistance related genes in a targeted manner; by utilizing the characteristics of nanopore length reading length and real-time sequencing and a tuberculosis specific targeted enrichment strategy, accurate identification of a mycobacterium tuberculosis complex group, synchronous typing of 42 mycobacteria, analysis of 24 drug-resistant genes, and efficient detection of structural variation and low-abundance heterogeneity drug-resistant mutation are realized, the detection period is shortened, the detection cost is reduced, and the detection efficiency is improved. And a comprehensive and reliable technical basis is provided for accurate diagnosis and treatment of mycobacterium infection.

The application relates to a mycobacterium tuberculosis drug resistance phenotype prediction method and device and a computer device. The method comprises the following steps: determining a to-be-predicted mutation site which does not exist in a mutation phenotype annotation database from at least one mutation site of a target gene fragment of to-be-processed mycobacterium tuberculosis according to gene sequencing data of the mycobacterium tuberculosis; converting protein structure change data of the to-be-predicted mutation site according to protein structure data corresponding to the to-be-predicted mutation site; inputting the protein structure change data of the to-be-predicted mutation site into a drug resistance phenotype prediction model to obtain a predicted drug resistance phenotype of the to-be-predicted mutation site; and determining a drug resistance phenotype prediction result of the to-be-processed mycobacterium tuberculosis based on the predicted drug resistance phenotype of the to-be-predicted mutation site and drug resistance phenotypes of the mutation sites except the to-be-predicted mutation site. The method can effectively improve the mycobacterium tuberculosis drug resistance phenotype prediction efficiency.

The invention relates to a mycobacterium tuberculosis EspB polyclonal antibody as well as a preparation method and application thereof. The purified EspB antibody has good detection specificity, only reacts with mycobacteria, and does not react with other common pneumonia pathogenic bacteria. The EspB polyclonal antibody specifically recognizes mycobacteria EspB, comprises mycobacterium tuberculosis and nontuberculous mycobacteria (NTM) EspB which grows slowly and rapidly, and can be used for detecting tuberculosis and NTM diseases.

The present invention relates to the use of a therapeutic agent based on cell wall fragments of a virulent strain of Mycobacterium tuberculosis-complex for the preparation of a drug for the treatment of patients with active tuberculosis.

The invention provides a mycobacterium tuberculosis drug sensitivity kit and a method and application thereof, the drug sensitivity condition of mycobacterium tuberculosis to 12 drugs such as bedaquiline is detected based on a liquid method, the kit comprises a drug freeze-drying agent and an additive, the drug freeze-drying agent contains drugs, glucan, glycerol and valine, and the additive is a mixture of the drugs, the glucan, the glycerol and the valine. The additive is used for culturing mycobacterium tuberculosis, acetic acid is added to a non-water-soluble drug as a solvent, a cosolvent or a disintegrating agent is added to an indissolvable drug, and the prepared drug freeze-drying agent can be stored for a long time, can be redissolved only with water, and is good in redissolution stability, free of solvent residual toxicity and higher in drug stability and drug sensitivity detection accuracy. And meanwhile, the method also has better universality.