63 results about "Pectobacterium" patented technology

The invention relates to a CRISPR detection primer set for MTBC (Mycobacterium Tuberculosis Complex) and use thereof, and belongs to the technical field of gene detection. The primer set includes an amplification primer pair and a crRNA; the amplification primer pair is used to amplify a common conserved sequence of M. tuberculosis, M. bovis, M. africanum, M. canettii, and M. microti; the crRNA includes an anchor sequence and a guide sequence, wherein the anchor sequence is bound to a cas protein, and the guide sequence is matched with a targeted RNA fragment in the common conserved sequence.Through the primer set based on CRISPR detection technology, the detection of common conserved sequences of M. tuberculosis, M. bovis, M. africanum, M. canettii, and M. microti by CRISPR detection canquickly perform on-site detection of MTBC, and have the advantages of high specificity, high sensitivity and simplicity.

The invention discloses a primer and a probe for on-site rapid detection of mycobacterium tuberculosis complex and a kit thereof. The forward primer sequence is shown as SEQ ID No.1, the reverse primer sequence is shown as SEQ ID No.2, and the probe sequence is shown as SEQ ID No.3. The invention provides the PA-LFD primer and the probe for on-site distinctive, sensitive, simple and rapid detection of the mycobacterium tuberculosis complex (mycobacterium tuberculosis, mycobacterium bovis, African mycobacteria and Canna mycobacteria) and the kit containing the primer and the probe.

The invention relates to a polymerase chain reaction (PCR) method for specifically detecting pectobacterium carotovorum. A pair of primers is designed according to a base sequence (GenBank:L32171.1) of a pectobacterium carotovorum subsp. carotovorum No.71 strain pectate lyase gene, and a basic local alignment search tool (BLAST) result proves that the pair of primers has high specificity. When the pair of primers is used for PCR detection, bacteria of other genera can be removed, the pectobacterium carotovorum can be distinguished from other bacteria of pectobacterium, and the specificity is high.

The present invention relates to diagnosis kit for Mycobacterium species identification and drug-resistance detection and manufacturing method thereof, which can discriminate a Mycobacterium Tuberculosis rpoB gene point mutation relating to the Mycobacterium species identification and drug-resistance swiftly, exactly and in large quantities using an oligonucleotide chip. The diagnosis kit for Mycobacterium species identification and drug-resistance detection in accordance with the present invention consists of an oligonucleotide chip including a Mycobacterium tuberculosis complex probe, a Mycobacterium species identification probe and a drug-resistance detection probe of a Mycobacterium tuberculosis rpoB gene, and a fluorescent material containing a biotin-binding protein so as to detect hybridization of amplified products of a specimen marked as biotine and the corresponding probe.

The invention discloses a primer sequence combination and a method for applying the primer sequence combination in PCR amplification. The amplification target in the method is specific fragments EMA inside the genome of Elizabethkingia meningoseptica, and the method shows excellent specificity and sensibility; and moreover, by using the method, the neisseria meningitidis, haemophilus influenzae, staphylococcus aureus, streptococcus pneumoniae, escherichia coli, listeria monocytogenes, mycoplasma pneumoniae, bordetella pertussis, klebsiella pneumoniae and mycobacterium tuberculosis and the other pathogenic bacteria which are likely to cause meningitis and diseases of the upper respiratory tract, and easy to be confused with the Elizabethkingia meningoseptica in an identification of bacteria, can be distinguished at one time. Additionally, the lower detection limit of the method can reach 10-4 ng per Mul.

Genes have been isolated from Pectobacterium cypripedii encoding geranylgeranyl pyrophosphate (GGPP) synthase (CrtE), phytoene synthase (CrtB), phytoene desaturase (Crtl), lycopene cyclase (CrtY), β-carotene hydroxylase (CrtZ), and zeaxanthin glucosyl transferase (CrtX) activity. The genes and their products are useful for the conversion of farnesyl pyrophosphate to carotenoids. Vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes by recombinant DNA technology in transformed host organisms are disclosed.

The invention relates to primers, probes and a method for liquid-phase chip detection of Mycobacterium tuberculosis drug-resistant gene mutation sites. The primers, probes and method are used for detecting drug-resistant gene mutation sites in Mycobacterium tuberculosis for drugs isoniazide, rifampicin and fonoquantel. The isoniazide drug-resistant mutation sites are positioned in katG gene and inhA gene; the rifampicin drug-resistant mutation sites are positioned in rpoB gene; and the fonoquantel drug-resistant mutation sites are positioned in gyrA gene. The method comprises the following steps: respectively carrying out homology analysis according to the nucleotide sequences of the four drug-resistance related genes in the gene bank, designing the primers and probes, carrying out PCR (polymerase chain reaction) twice, carrying out molecular hybridization, and carrying out detection by using a Luminex200 system, thereby determining whether the sample contains the drug-resistant mutation sites. The detection of drug-resistant gene mutation sites is of crucial importance for treating Mycobacterium tuberculosis infection by adopting correct therapeutic schedules. The primers, probes and method have the advantages of high detection speed, high sensitivity, high specificity and the like, are simple to operate, and are beneficial to popularization and application.

The invention relates to a method for extracting live bacteria RNA in Mycobacterium tuberculosis and a detection kit thereof, and particularly provides a method for extracting live bacteria RNA in Mycobacterium tuberculosis and a detection kit which is used for the live bacteria RNA in the Mycobacterium tuberculosis and is obtained by applying the method and combining fluorescent quantitative PCR technology. The kit can accurately, sensitively and quickly identify dead bacteria and the live bacteria of the Mycobacterium tuberculosis, reduce the cost and shorten the detection time, and more importantly, the kit is the basis of studies such as the diagnosis of tuberculosis, medicinal susceptibility experiments, the monitoring of chemotherapy response, the screening of new antitubercular medicaments, the prevention of the tuberculosis and the like.

The invention discloses a DNA marker for detecting the drug resistance of mycobacterium tuberculosis and application of the DNA marker. It is found that mutant protein Rv2783cD67N has the DNA polymerase activity without dependence on a DNA template, is not suppressed by POA and has the phosphorolysis singe-stranded DNA (SS DNS) activity; besides, the mutant protein Rv2783cD67N can not be combined with POA; by means of the locus mutation, the drug resistance of mycobacterium tuberculosis on PZA can be remarkably caused, the locus can serve as a mark locus for molecular diagnosis of PZA drug resistance of mycobacterium tuberculosis, and the detection rate and the precision of mycobacterium tuberculosis with PZA drug resistance can be easily improved. The diagnosis time of patients can be effectively shortened, and the treatment time and cost of patients can be saved. The DNA marker for detecting the drug resistance of mycobacterium tuberculosis and application of the DNA marker provide a new thought for finding new and more effective tuberculosis drug resistance molecular markers.

The invention discloses a preparation method and application of a series of novel pyridine derivatives. The derivatives can be used for the treatment of related diseases caused by mycobacterium species, especially for diseases caused by pathogenic mycobacterium, such as Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium avium and Mycobacterium marinum.

The invention relates to the field of agricultural microorganisms, and discloses a beauveria bassiana and an application thereof. The preservation number of the beauveria bassiana disclosed by the invention is CGMCC (China General Microbiological Culture Collection Center) No. 21047. In addition, the invention also provides an application of the beauveria bassiana in preparation of a hydroxamic acid type siderophore and regulation of plant diseases. The beauveria bassiana disclosed by the invention is a dominant strain of high-yield siderophore, and can be used for synthesizing and secreting the hydroxamic acid type siderophore. The secondary metabolites of the beauveria bassiana disclosed by the invention have antibacterial activity on pseudomonas solanacearum, pectobacterium carotovorumand ralstonia solanacearum.

The invention discloses Lactobacillus casei WX322 with a preservation number of CGMCC No.17710, a strain and its metabolites have broad-spectrum antibacterial activity, which can exhibit strong antibacterial ability against a variety of Gram-positive and Gram-negative bacteria (including pathogenic and spoilage bacteria), can be used to prepare antibacterial or / and antiseptic products, and can beused in the fields of medical, food and other industry to control bacterial pathogens or spoilage bacteria, can also effectively inhibit the growth of the soft rot pathogen Pectobacterium carotovorum,and can be used to prepare products for controlling bacterial soft rot of vegetables, can control the decay of vegetables caused by Pectobacterium carotovorum before and after harvest, can effectively solve the problem of agricultural residues and environmental pollution caused by pesticide control, has good application potential in the biological control of vegetable bacterial soft rot, and alsoprovides an excellent basic strain for the development of microbial pesticides.

The invention provides a primer combination and an application thereof. The primer combination comprises a first primer group and a second primer group, wherein the nucleotide sequence of the first primer group is shown in the SEQ ID NO:1-2, and the nucleotide sequence of the second primer group is shown in the SEQ ID NO:3-4. The primer combination can specifically recognize the mycobacteria specific area-ITS area, has high homology between different mycobacteria, and is applicable to various mycobacteria and capable of being effectively used for recognizing the mycobacteria, particularly tuberculous mycobacteria and non-tuberculous mycobacteria.

The present invention provides a kit for rapid detection of Mycobacterium tuberculosis in a sputum sample, and the kit includes a liquefied solution, a lysis solution, a probe solution, a stop solution, a fixative solution, a carrying glass sheet and a cover glass sheet. After liquefaction of the sputum sample, cell lysis, probe hybridization and fluorescence fixing and other operations are performed, and a fluorescence microscopy is used for macroscopic observation, counting and analysis of the Mycobacterium tuberculosis. The kit is simple in operation method, short in procedure, easy to operate, strong in test result specificity, high in sensitivity, clear in results and high in reliability, and can be used for direct, simple and effective detection of the Mycobacterium tuberculosis in the sputum sample.

The invention discloses a method for identifying mycobacterium. The method comprises the following steps: acquiring a plurality of standard sequences of a plurality of mycobacteria so as to generate a mycobacterium sequence database; by comparing the standard sequences, obtaining a conservation section mutually owned by the standard sequences; preparing a PCR primer according to the conservation section; using the PCR primer to perform the PCR amplification of a DNA solution of the testing mycobacterium so as to obtain an amplification product of the testing mycobacterium; carrying out the sequencing reaction of the amplification product to obtain the sequencing product; carrying out the sequencing analysis of the sequencing product to obtain a sequence of the testing mycobacterium; and by comparing the testing sequence with the standard sequence in the mycobacterium sequence database, determining the type of the testing mycobacterium. The PCP primer is designed according to the conservation section of the standard sequences of various mycobacteria, only a pair of primers is needed for all PCRs and sequencing, thereby greatly reducing the quantity of oligonucleotide needed for experimental population.

The present invention discloses a PCR-RLB assay process of mycobacteria and the employed primers and probes. A pair of oligonucleotide primers and 35 species or type-specific probes are designed against the characteristics of DNA sequences of different types of mycobacterias. Via PCR-RLB assay process, 17 mycobacterias and Mycobacterium tuberculosis complexes, Mycobacterium Ulcerans / Marinum and 2 subspecies of Mycobacterium Kansasii can be identified for SGM. 14 species or subspecies can be identified for RGM. Compared with traditional identification process of mycobacteria, the present invention is a rapid, simple, practical, specific and sensitive assay method.

The invention discloses a primer pair for detecting and identifying Pectobacterium bacteria and a using method thereof. The primer comprises a forward primer 23SPecF and a reverse primer 23SPecR. The method for detecting and identifying the Pectobacterium bacteria comprises the following steps of: performing Polymerase Chain Reaction (PCR) on the 23SPecF and the 23SPecR by using the primer pair, wherein a 266bp nucleic acid fragment can be only amplified from a sample containing the genome DNA of the Pectobacterium bacteria. Due to that the Pectobacterium bacteria have seven copied 23SrRNA genes, the detection sensitivity of the primer pair for performing the PCR on the 23SPecF and the 23SPecR is seven times that of a single-copy gene primer, and the Pectobacterium bacteria can be detected more sensitively.

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 method for converting plant sterols into 9 alpha-hydroxyandrostenedione by using a Mycobacterium fortuitum ATCC 35855 bacterial strain. Especially the method uses ultrasonic wave vibration.

The present invention relates to a chimeric DNA vaccine for preventing tuberculosis and immunotherapy, comprising a full-length open reading frame of the nucleotide sequence shown in SEQ ID NO.1, or a conservative variant thereof, in SEQ ID NO.1 It comprises mycobacterium hsp70 gene and human B7-1 gene, and a linker sequence is designed between the two genes to connect them. The vaccine of the invention can be used for tuberculosis caused by Mycobacterium tuberculosis; or express the corresponding protein in eukaryotic cells in vitro for preventing and treating tuberculosis. It can also be used as an immune enhancer for corresponding viral diseases and tumor immunotherapy.

The present invention relates to diagnosis kit for Mycobacterium species identification and drug-resistance detection and manufacturing method thereof, which can discriminate a Mycobacterium Tuberculosis rpoB gene point mutation relating to the Mycobacterium species identification and drug-resistance swiftly, exactly and in large quantities using an oligonucleotide chip. The diagnosis kit for Mycobacterium species identification and drug-resistance detection in accordance with the present invention consists of an olignucleotide chip including a Mycobacterium tuberculosis Mycobacterium species identification probe and a drug-resistance detection probe of a complex probe, a Mycobacterium tuberculosis rpoB gene, and a fluorescent material containing a biotin-binding protein so as to detect hybridization of amplified products of a speciment marked as biotine and the corresponding probe.

This invention discloses a method for identifying multiple PCR primers of tuberculous mycobacteria strains, which comprises the steps of: (1) adding an abacerial genome sequence to 5' end of oligonucleotide primers P1 and P2 that can combine with the bacterium genome sequence to obtain specific long primers YB1-P1and YB2-P2 as the primer pair for the polymerase chain reaction in the first stage of multiplex amplification process; and (2) directly using the abacterial genome sequence as the peimer pair for the polymerase chain reaction in the first stage of multiplex amplification process.

The invention discloses a primer and a probe for on-site rapid detection of mycobacterium tuberculosis complex and a kit thereof. The forward primer sequence is shown as SEQ ID No.1, the reverse primer sequence is shown as SEQ ID No.2, and the probe sequence is shown as SEQ ID No.3. The invention provides the PA-LFD primer and the probe for on-site distinctive, sensitive, simple and rapid detection of the mycobacterium tuberculosis complex (mycobacterium tuberculosis, mycobacterium bovis, African mycobacteria and Canna mycobacteria) and the kit containing the primer and the probe.

The present invention relates to the use of a strain of a mycobacterium of the Mycobacterium tuberculosis complex, into which an mspA gene capable of expressing a Mycobacterium smegmatis porin A has been inserted, for producing a vaccine for the prevention of an infection by a bacterium of the Mycobacterium tuberculosis complex in a host comprising eukaryotic cells, preferably macrophages, wherein said mycobacterial strain thus transformed exhibits reduced growth in said eukaryotic cells.

The invention discloses an N-acylhomoserinelactone degrading enzyme and application thereof. The degrading enzyme contains a gene AhlM, and the nucleotide sequence of the AhlM is SEQ NO. 1. The N-acylhomoserinelactone degrading enzyme containing the gene AhlM can degrade AHL generated by pathogenic bacteria Pectobacterium spp, Pec, is good in heat stability and pH stability and can obviously inhibit soft rot of crops caused by the pathogenic bacteria Pec.

The invention belongs to the technical field of microbial application. According to a biocontrol agent for controlling pectobacterium atroseptica, the content of viable bacilli is larger than or equalto 1 * 108 cfu per gram of bacterial agent, and the bacilli are composite bacilli of bacillus pumilus, bacillus megaterium, bacillus amyloliquefaciens, bacillus licheniformis and bacillus mucilaginosus. The biocontrol agent has good biocontrol and growth promoting effects, and can effectively control pectobacterium atroseptica.

The invention discloses an RPA (Recombinase Polymerase Amplification) primer, kit and fast detecting method for fast detecting potato black leg diseases (pectobacterium spp.), and relates to the fieldof plant disease detection. The RPA primer has nucleotide sequences shown as SEQ ID NO.1-2. The potato black leg diseases (pectobacterium spp.) can be detected by only performing isothermal amplification on the extracted DNA of potato black leg disease (pectobacterium spp.) bacteria for 20 min by using the RPA primer. The RPA detection primer of the potato black leg diseases (pectobacterium spp.)has the advantages that the specificity is high; the sensitivity is high; the detection is efficient and fast; the operation is simple and convenient; a special instrument is not needed; and the technical basis can be provided for early diagnosis of the potato black leg diseases (pectobacterium spp.).

The invention discloses a method to rapidly check out the nodule embranchment bacilli and the non-nodule embranchment bacilli. It includes the following steps: under the condition of amplifying specificity, taking polymerase chain reaction in polymerase chain system. The invention supplies the corresponding agent box. It could rapidly and easily check out and identify the nodule embranchment bacilli and the non-nodule embranchment bacilli.

The invention relates to an isolated DNA construct, comprising a mycobacterium secreted acid phosphatase promoter or promoter fragment which is inducible by low pH. This invention further relates to diagnostic methods and vaccines for treatment or prophylaxis of pathogenic mycobacterial disease or infection. This invention also provides for methods of screening for compounds capable of modulatingthe activity, production or secretion of a mycobacterium secreted acid phosphatase.