1006 results about "Mycobacterium" patented technology

The invention provides recombinant escherichia coli with high yield of ergothioneine and a method for preparing ergothioneine, the recombinant escherichia coli co-expresses ergothioneine anabolic enzymes Egt1 and Egt2, the methyltransferase structural domain of the Egt1 is derived from trichoderma reesei, mycobacterium smegmatis EgtD or rhizopus nigricans, and the methyltransferase structural domain of the Egt2 is derived from the trichoderma reesei, the mycobacterium smegmatis EgtD or the rhizopus nigricans. The histidine sulfoxide synthase structural domain is derived from trichoderma reesei, schizosaccharomyces pombe or rhizopus nigricans, and the Egt2 is derived from claviceps purpurea, neurospora crassa or aspergillus niger. After the recombinant escherichia coli obtained by screening is cultured for 68 hours in a 5L fermentation tank, the yield of ergothioneine can reach 5g / L or above; wherein after recombinant escherichia coli co-expressing Egt1 enzyme (the structural domain of methyltransferase is derived from mycobacterium smegmatis EgtD, and the structural domain of histidine sulfoxide synthase is derived from trichoderma reesei) and Egt2 enzyme (derived from neurospora crassa) are cultured in a 5L fermentation tank for 68 hours, the yield of ergothioneine can reach 8.7 g / L, and the production intensity reaches 127.9 mg / (L.h).

Provided herein are methods for treating a pulmonary infection in a patient in need thereof, for example, a nontuberculous mycobacterial pulmonary infection for at least one treatment cycle. The method comprises administering to the lungs of the patient a pharmaceutical composition comprising a liposomal complexed aminoglycoside comprising a lipid component comprising electrically neutral lipids and an aminoglycoside. Administration comprises aerosolizing the pharmaceutical composition to provide an aerosolized pharmaceutical composition comprising a mixture of free aminoglycoside and liposomal complexed aminoglycoside, and administering the aerosolized pharmaceutical composition via a nebulizer to the lungs of the patient. The methods provided herein result in a change from baseline on the semi-quantitative scale for mycobacterial culture for a treated patient, and / or NTM culture conversion to negative during or after the administration period.

Provided herein are methods for treating a pulmonary infection in a patient in need thereof, for example, a nontuberculous mycobacterial pulmonary infection for at least one treatment cycle. The method comprises administering to the lungs of the patient a pharmaceutical composition comprising a liposomal complexed aminoglycoside comprising a lipid component comprising electrically neutral lipids and an aminoglycoside. Administration comprises aerosolizing the pharmaceutical composition to provide an aerosolized pharmaceutical composition comprising a mixture of free aminoglycoside and liposomal complexed aminoglycoside, and administering the aerosolized pharmaceutical composition via a nebulizer to the lungs of the patient. The methods provided herein result in a change from baseline on the semi-quantitative scale for mycobacterial culture for a treated patient, and / or NTM culture conversion to negative during or after the administration period.

The invention belongs to the technical field of probiotics and application thereof, and relates to a probiotic capable of reducing uric acid and a product thereof, in particular to a strain of Lentilactobacillusbuchneri strain ZD62 which is preserved in China Center for Type Culture Collection on March 14, 2024, the preservation address is Wuhan University, Wuhan, China, and the preservation number is CCTCC NO: M2024469, and the preservation number is CCTCC NO: M2024469, the preservation number is CCTCC NO: M2024469, the preservation number is CCTCC NO: M2024469, and the preservation number is CCTCC NO: M2024469. The invention relates to a compound probiotic agent, which is prepared from Lentilactobacilli buchneri strain ZD62 and Mycobacterium fermentum ZD103, and is characterized in that the Mycobacterium fermentum ZD103 is prepared from Lentilactobacilli buchneri strain ZD62; the lactobacillus buchneri ZD62 and the mucus fermentum ZD103 provided by the invention have a good characteristic of reducing uric acid.

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.

Provided herein are systems for treating a subject with a pulmonary infection, for example, a nontuberculous mycobacterial pulmonary infection, a Burkholderia pulmonary infection, a pulmonary infection associated with bronchiectasis, or a Pseudomonas pulmonary infection. The system includes a pharmaceutical formulation comprising a liposomal aminoglycoside dispersion, and the lipid component of the liposomes consist essentially of electrically neutral lipids. The system also includes a nebulizer which generates an aerosol of the pharmaceutical formulation at a rate greater than about 0.53 gram per minute. The aerosol is delivered to the subject via inhalation for the treatment of the pulmonary infection.

The invention relates to the technical field of high-throughput targeted sequencing, in particular to a primer group for detecting important drug-resistant genes and virulence genes of multi-drug-resistant pathogenic bacteria and application of the primer group. Specifically, the kit comprises detection primers for multiple drug-resistant bacteria such as third-generation cephalosporin drug-resistant enterobacteriaceae bacteria, carbapenem drug-resistant enterobacteriaceae bacteria, carbapenem drug-resistant acinetobacter baumannii, rifampicin drug-resistant mycobacterium tuberculosis and the like. The method has the technical advantages of rapidness, high efficiency, strong targeting property, strong specificity and the like, is suitable for epidemic and difference analysis of various important drug-resistant genes and virulence genes carried by metagenome samples of various environments or disease materials and the like, evaluates the risk of related environments on the basis, and can further guide prevention and clinical treatment medication.

The invention belongs to the technical field of biology, and relates to a recombinant mycobacteriophage capable of enabling host bacteria to express nano luciferase as well as a preparation method and application of the recombinant mycobacteriophage. The recombinant mycobacterium fluorescent bacteriophage capable of enabling the host bacteria to express the nano luciferase comprises an expression element for expressing the nano luciferase in mycobacteria, the expression element contains a gene nluc required by luminescence and a mycobacterium strong promoter, and the upstream of the gene nluc is connected with a promoter sequence to ensure the normal expression of downstream genes. The base sequence is as shown in SEQ ID NO: 1. The preparation method comprises the following steps: carrying out enzyme digestion on a bacteriophage skeleton to form a linear sequence, enabling the linear sequence to be recombined with a target fragment containing a gene nulc, connecting the enzyme-digested bacteriophage skeleton with the target fragment containing the gene nluc, carrying out in-vitro packaging, transferring into an escherichia coli competent cell, and carrying out in-vivo amplification in escherichia coli; the method comprises the following steps: extracting positive phagocytids of recombinant escherichia coli, electrically transferring the phagocytids into Mycobacterium smegmatis, and culturing the phagocytids on a double-layer agar plate to obtain the bacteriophage. According to the application of the recombinant mycobacterium bacteriophage in the mycobacterium rapid detection and / or drug sensitivity detection reagent, the method is simple, convenient and rapid, the sensitivity is high, and the repeatability is strong.

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 two mycobacterium bacteriophages and application thereof. The two bacteriophages both have a splitting spectrum width and efficient splitting capability, can specifically realize effective splitting of mycobacteria of five specific sequence types, and are further applied to the field of prevention and control of mycobacteria. Specifically, the bacteriophage MP2435 and the bacteriophage MP2451 not only can effectively inhibit the proliferation process of mycobacteria and block the formation of a biofilm of the mycobacteria, but also can efficiently kill the mycobacteria in a milk matrix; meanwhile, the two bacteriophages can still keep relatively high activity under the conditions that the temperature is 37-65 DEG C and the pH value is 3-12, and the bactericidal activity of the bacteriophages can be stably maintained even in acid-base, high-temperature and ultraviolet-containing extreme environments. Through the characteristics, the application scene and application range of the bacteriophage in mycobacterium prevention and control are remarkably widened, and an efficient and stable novel technical means is provided for biological prevention and control of mycobacterium.

The invention provides a genetically engineered bacterium for high yield of 9 alpha-hydroxyandrostane-4-ene-3, 17-diketone as well as a construction method and application of the genetically engineered bacterium for high yield of 9 alpha-hydroxyandrostane-4-ene-3, 17-diketone. According to the invention, the production of by-products 4-HBC, 4-AD and 9-OH-3-OPCM is eliminated by blocking the expression of Opccr, SalA and TeB in mycobacterium neoaurum and overexpressing KshA1, the production limitation of 9-OHAD is liberated by overexpressing rate-limiting enzyme ChsH1-2, and finally, under the condition that the concentration of phytosterol is 20g / L, the purposes that the yield of 9-OHAD is 13.4 g / L, the molar yield is 91.1% and almost no by-product exists are achieved. When amplification is carried out at the concentration of 45g / L of phytosterol in a 5L bioreactor, soybean oil and (2-hydroxypropyl)-beta-cyclodextrin are used as auxiliaries, and a 9-OHAD conversion result at the concentration of 20g / L in a shake flask is basically reproduced.

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 provides a primer group and a kit for detecting bovine mycobacterium paratuberculosis based on a recombinase isothermal amplification technology and application, and relates to the technical field of biological detection.The primer group comprises a forward primer MAP-F3, a reverse primer MAP-R3 and a specific fluorescent probe MAP-P3 and has good specificity; the sensitivity reaches 10fg / mu L, which is more sensitive than that of a colloidal gold test strip, the dependence on equipment is broken through, and the detection result is more accurate. The technical problem that in the prior art, it is difficult to distinguish bovine mycobacterium paratuberculosis from other pathogens through a detection method, or the detection process is tedious is solved. According to the prepared reagent or kit, the liquid preparation process is simple, the whole amplification process can be completed only in 30 min, the amplification yield can reach 109-1010 copies, the method is rapid and efficient, the amplification result can be directly judged according to fluorescence data, other auxiliary means are not needed, and the method is suitable for field detection.

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 invention discloses a traditional Chinese medicine composition for resisting giant frog blue eye disease pathogen Elizabeth mileli and application of the traditional Chinese medicine composition, and relates to the technical field of aquaculture disease prevention and treatment. It is found for the first time that the radix scutellariae has a good effect of inhibiting the Elizabeth mille, and the herba houttuyniae does not have the effect of inhibiting the Elizabeth mille but can enhance the inhibiting effect of the radix scutellariae on the Elizabeth mille, that is, the herba houttuyniae has a synergistic enhancement effect; the traditional Chinese medicine composition of the radix scutellariae and the herba houttuyniae is expected to be applied to treatment of the blue eye disease of the giant spiny frog, and the Chinese herbal medicine composition is used for replacing antibiotics in the breeding process.

The invention discloses application of nadifloxacin, and relates to the technical field of biological medicines. According to the application disclosed by the invention, the condition that the nadifloxacin not only can better inhibit the Mycobacterium tuberculosis, but also can better inhibit the Mycobacterium nontuberculosis is found for the first time, so that the nadifloxacin can be used for preparing an inhibitor of the Mycobacterium tuberculosis and / or the Mycobacterium nontuberculosis; the invention also relates to a medicine for treating diseases caused by dry infection of Mycobacterium tuberculosis and / or non-Mycobacterium tuberculosis.

The invention provides a preparation method and application of nano protein, and the preparation method comprises the following steps: adopting lactoferrin as a carrier, reducing by using a reducing agent, stirring with DNA enzyme and a DNA chimeric agent, and obtaining the nano protein by using a nano precipitation method, the preparation method comprises the following steps: by taking lactoferrin as a nano-carrier, opening a folding structure of the lactoferrin by utilizing glutathione, then adding chloroquine and DNA enzyme, and finally, accelerating the reformation of a disulfide bond of the lactoferrin by stirring. On one hand, a DNA chimeric agent chloroquine is utilized to maintain the eDNA configuration in the biological membrane to be B type, and on the other hand, the mycobacterium tuberculosis biological membrane is efficiently disintegrated based on the enzymatic hydrolysis capability of DNA enzyme; the nano protein can efficiently grab iron ions at a focus part based on the iron chelation characteristic of the nano protein, so that bacteria die due to iron deficiency, and the bioavailability of DNA enzyme and the hydrolysis efficiency of a biological membrane can be improved to a great extent.

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.

The invention discloses a mycobacterium tuberculosis tandem DNA vaccine W543 as well as a preparation method and application thereof. The DNA vaccine W543 provided by the invention is a novel mycobacterium tuberculosis DNA vaccine with multiple antigen immunodominance epitopes, which is constructed by connecting polypeptides or epitope coding genes of a mycobacterium tuberculosis proliferation period antigen and a latent related antigen in series through a linker by virtue of genetic engineering and inserting the polypeptides or epitope coding genes into an eukaryotic expression vector pVAX1. Experiments prove that the mycobacterium tuberculosis specific tandem DNA vaccine W543 disclosed by the invention is used as an immunomodulatory preparation, mainly stimulates immune response of Th1 type, and can significantly enhance the specific cellular immune function of mice; as a preventive treatment preparation, the bacterial count of lung and liver tissues can be reduced, the lesion range of the lung tissues is remarkably reduced, the lesion is relieved, and the treatment effect is slightly superior to that of an existing commercial tuberculosis latent infection prevention and treatment preparation, namely, microkalidium vaccine.

The present invention relates to a method of achieving a positive therapeutic response against nontuberculous mycobacterial (NTM) infections by administering an oral dose of epetravorole to a human subject.

The invention provides a method for improving cell permeability of mycobacterium neoaurum, a mycobacterium neoaurum engineering bacterium and an application of the mycobacterium neoaurum engineering bacterium in preparation of 9 alpha-hydroxyandrostane-4-ene-3, 17-diketone (9-OHAD). Biotransformation of phytosterol into high-value steroid intermediates such as 9-OHAD in mycobacteria is a footstone of steroid drugs. However, the limited permeability of its dense cell wall of Mycobacterium severely hinders the effective intracellular transport of phytosterols and its bioconversion to 9-OHAD. The endogenous synthesis capability and exogenous circulation capability of mycobacterium neoaurum trehalose are destroyed through gene knockout, the permeability of cells is improved, the molar conversion rate of 9-OHAD is increased to 65.7% from 44.7%, the extracellular production of 9-OHAD is also improved, the extracellular yield of 9-OHAD is increased to 32.1% from 12%, and subsequent separation and purification are facilitated.

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.

Provided herein are methods for treating a pulmonary infection in a patient in need thereof, for example, a nontuberculous mycobacterial pulmonary infection for at least one treatment cycle. The method comprises administering to the lungs of the patient a pharmaceutical composition comprising a liposomal complexed aminoglycoside comprising a lipid component comprising electrically neutral lipids and an aminoglycoside. Administration comprises aerosolizing the pharmaceutical composition to provide an aerosolized pharmaceutical composition comprising a mixture of free aminoglycoside and liposomal complexed aminoglycoside, and administering the aerosolized pharmaceutical composition via a nebulizer to the lungs of the patient. The methods provided herein result in a change from baseline on the semi-quantitative scale for mycobacterial culture for a treated patient, and / or NTM culture conversion to negative during or after the administration period.

The invention relates to the technical field of gene detection, and discloses a mycobacterium tuberculosis drug resistance gene detection data analysis system which comprises a data import module, a signal correction module, a feature extraction module, a multi-stage interpretation module, a result fusion module and a report generation module. The system performs baseline drift correction, abnormal point detection and nonlinear normalization on a fluorescence intensity value output by a detector, extracts multi-dimensional features such as peak intensity, a mutation ratio, a consistency coefficient and an in-batch difference index, and constructs a dynamic threshold function to realize wild type and mutation type layered judgment. A drug resistance risk conclusion is generated through drug resistance characteristic matrix mapping, a standardized detection report is output, and intelligent analysis of a detection result and automatic report generation are achieved. Through multi-stage signal correction, multi-dimensional feature extraction and dynamic threshold interpretation, intelligent analysis and standardized report output of a mycobacterium tuberculosis drug resistance gene detection result are realized, and data judgment accuracy and clinical application efficiency are improved.

The invention provides a composition and a kit for detecting mycobacterium tuberculosis and drug resistance thereof. Specifically, the invention provides a composition of primers and probes for detecting mycobacterium tuberculosis and drug resistance thereof, and genes targeted by the primers and the probes comprise an IS6110 insertion sequence, an IS1081 insertion sequence, a 16s rRNA sequence, an rpoB gene, a katG gene, an inhA gene, an ahpC gene, a gyrA gene and a gyrB gene.

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 relates to novel rufomycin analogs targeting C1pC1 protease. The compounds of the invention have antibacterial activity, for example, with anti-tuberculosis properties, and are useful for the treatment of, for example, 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 belongs to the field of biological detection, and discloses an efficient and rapid concentration method and device for LPS or LAM. The inventor finds out that the non-polar macroporous resin with a specific aperture range has similar affinity adsorption efficiency to lipopolysaccharide or lipoarabinomannan, the adsorption capacity is large, the adsorption speed is high, the lipopolysaccharide can be efficiently eluted in cooperation with a specific eluent, a concentrated lipopolysaccharide to-be-detected sample is obtained, and the non-polar macroporous resin with the specific aperture range can be used for detecting the lipopolysaccharide or lipoarabinomannan in the lipopolysaccharide or lipoarabinomannan in the lipopolysaccharide or lipoarabinomannan. And a good condition is created for the detection of lipopolysaccharide or lipoarabinomannan. Efficient concentration of LAM is realized by using the method, and excellent conditions are provided for detection of mycobacterium tuberculosis.