43 results about "Streptococcus dysgalactiae" patented technology

Disclosed are antibodies that block the binding of fibronectin protein to fibronectin. Also disclosed are site specifically-mutated and truncated peptide epitopes derived from the fnbA and fnbB genes of Staphylococcus aureus, the fnba and fnbB genes of Streptococcus dysgalactiae, and the sfb gene of Streptococcus pyogenes, and nucleic acid segments encoding these peptides and epitopes. The anti-(fibronectin binding site) antibodies, peptides and epitopes that give rise to antibodies that block the binding of fibronectin binding proteins to fibronectin, and DNA segments encoding these proteins and are of use in various screening, diagnostic and therapeutic applications including active and passive immunization and methods for the prevention of streptococcal and staphylococcal colonization in animals or humans. These. DNA segments and the peptides derived therefrom are proposed to be of use directly in the preparation of vaccines and also for use as carrier proteins in vaccine formulations.

The invention provides a triple real-time fluorescent PCR method and a kit for detecting three streptococci at the same time. The kit comprises: a streptococcus agalactiae detection primer pair which is represented as the Seq ID No.1 and the Seq IN No.2 in the sequence table, a probe which is represented as the Seq ID No.3 in the sequence table, a streptococcus dysgalactiae detection primer pair which is represented as the Seq ID No.4 and the Seq ID No.5 in the sequence table, a probe which is represented as the Seq ID No.6 in the sequence table, a streptococcus iniae detection primer pair which is represented as the Seq ID No7 and the Seq ID No.8 in the sequence table, a probe which is represented as the Seq ID No.9 in the sequence table, three positive comparison sample which are represented as the Seq ID No.16, the Seq ID No.17 and the Seq ID No.18 in the sequence table, a negative comparison sample (ddH2O), a PCR buffering liquid required in fluorescent real-time quantification PCR amplification, dNTP and TaqDNAPolymerase. The method is simple and quick and is good in specificity.

The invention relates to the technical field of gene engineering, in particular to novel broad-spectrum chimeric lysin BGS-PlySb and an encoding gene and application thereof. The novel broad-spectrum chimeric lysin BGS-PlySb has an amino acid sequence shown as in SEQ ID NO. 1; the encoding gene of the novel broad-spectrum chimeric lysin BGS-PlySb has a nucleotide sequence shown as in SEQ ID NO. 2. A novel chimeric lysin is constructed by means of gene splicing and is suitable for killing various species of Streptococcus and Staphylococcus, particularly various species of Enterococcus, and Staphylococcus aureus, as well as Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus suis, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus mutans, Streptococcus iniae and other species; the novel broad-spectrum chimeric lysin BGS-PlySb has good stability and is insensitive to high-concentration NaCl, recombinant protein GBS-PlySb is well expressible in Escherichia coli BL21 (DE3), and a high dose of GBS-PlySb is free of significant cytotoxicity. Therefore, the novel broad-spectrum chimeric lysin BGS-PlySb is applicable independently to or as an additive to the control of various species of Streptococcus and the treatment of infections caused by these species, and has a promising application prospect.

The invention discloses an LAMP primer combination for detecting 8 environmental pathogens of dairy cow mastitis and application thereof. The primer combination provided by the invention is composed of 48 primers shown in sequence 1 to sequence 48. The primer combination provided by the invention can be used for detecting whether a to-be-detected bacterium is Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus dysgalactiae, Streptococcus uberis, Salmonella typhimurium, Proteus mirabilis or Candida albicans, and can be used for detecting whether a to-be-detected sample contains Escherichia coli and / or Klebsiella pneumonia and / or Pseudomonas aeruginosa and / or Streptococcus dysgalactiae and / or Streptococcus uberis and / or Salmonella typhimurium and / or Proteus mirabilis and / or Candida albicans. The primer combination provided by the invention is used for identifying and detecting the 8 environmental pathogens of dairy cow mastitis, has high specificity and high sensitivity, and can realize simple, rapid and accurate detection, thus having great popularization value.

The invention discloses a culture medium for the growth of bovine-derived streptococcus agalactiae or streptococcus dysgalactiae. The components of the bovine-derived streptococcus agalactiae culture medium are as follows: 1000ml of beef extract, 17g to 23g of tryptone, 2g to 5g of yeast extract powder, 2g to 5g of sodium chloride, 0.3g to 0.5g of disodium hydrogen phosphate, 2g to 3g of sodium bicarbonate, 50ml to 60ml of whey and 20ml to 25ml of glucose solution by 500g / L. The components of the bovine-derived streptococcus dysgalactiae culture medium are as follows: 1000ml of beef extract, 17g to 23g of tryptone, 2g to 5g of yeast extract powder, 2g to 5g of sodium chloride, 0.3g to 0.5g of disodium hydrogen phosphate, 2g to 3g of sodium bicarbonate, 20ml to 25ml of calf serum, 50ml to 60ml of whey and 20ml to 25ml of glucose solution by 500g / L.

This invention discloses a lysin that can kill many species of Streptococci. A new lysin, ClyR, was constructed by the gene splicing method. The ClyR can effectively kill different species of Streptococci, including Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus suis, Streptococcus uberis, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus mutans, Streptococcus equi, and various Enterococci and Staphylococcus aureus. ClyR shows good stability and is not sensitive to EDTA and high concentration of NaCl. Moreover, the ClyR is active in a wide range of pH and maintains high activity in pH 5-11. Recombinant protein ClyR is well expressed in E. coli stain BL21 (DE3). High doses of ClyR showed no apparent toxicity in mice. Furthermore, administration of 0.8 mg per mouse once is able to completely protect the mouse infected with lethal doses of Group B Streptococci. The ClyR can be used alone or in combination with different forms of reagents and solutions, for the control of a variety of Streptococci and for the treatment of infections caused by these bacteria. It has a broad application prospect.

Provided is a Streptococcus dysgalactiae ID9103 strain having accession number KCTC11818BP, and a method of producing hyaluronic acid by culturing the strain to produce hyaluronic acid having an average molecular weight of 10,000,000 Da or more.

The invention provides a GIT fusion protein used for preventing dairy cow mastitis. The GIT fusion protein has an amino acid sequence as represented by SEQ ID No. 1 or an amino acid sequence which is formed after substitution, deletion or addition of one or a plurality of amino acids of the sequence represented by SEQ ID No. 1 and has same functions as the sequence represented by SEQ ID No. 1 does. The invention also provides the nucleotide sequence of the protein and a preparation method and application for the same. The protein has good immunogenicity, can effectively prevent infection caused by Staphylococcus aureus, Streptococcus dysgalactiae, Streptococcus agalactiae and Streptococcus uberis, has a wide immune range and can be used for preventing dairy cow mastitis; moreover, while immunoprotection effects of the GIT fusion protein provided in the invention is guaranteed, preparation process and immune procedures in actual production are simplified, the length of the peptide chain of the fusion protein is shortened, expression of the fusion protein is enhanced, and the GIT fusion protein has an important value in development and application of novel vaccines.

The invention discloses streptococcus and a dairy cow mastitis vaccine obtained by inactivating the streptococcus. The active components of the dairy cow mastitis vaccine are an inactivated strain SAWR-6 (streptococcus agalactiae-6), an inactivated strain SDTR-9 (streptococcus dysgalactiae-9) and an inactivated strain SURF-5 (streptococcus uberis-5). The invention also provides a dairy cow mastitis vaccine, of which the active components are inactivated strains and capsular polysaccharides, wherein the inactivated strains are an inactivated strain SAWR-6, an inactivated strain SDTR-9 and an inactivated strain SURF-5; the capsular polysaccharides are strain SAWR-6 capsular polysaccharide, strain SDTR-9 capsular polysaccharide and strain SURF-5 capsular polysaccharide. The vaccine disclosed by the invention has the advantages of high efficiency, universality, safety, stability, various preparations and the like, a powerful guarantee is provided to safe, effective and comprehensive and effective prevention of streptococcus infected dairy cow mastitis, and the vaccine is a new milestone for the development of dairy cow mastitis vaccine.

The invention discloses a pharmaceutical composition for treating bacterial dairy cow mastitis. The pharmaceutical composition is prepared from the following raw materials: N-acetylcysteine, ampicillin, polyethylene glycol 400 and polyethylene glycol 4000; the invention further provides a preparation method of the pharmaceutical composition. The pharmaceutical composition has the beneficial effects that a preparation process is simple; the pharmaceutical composition is convenient to use in clinic; after the N-acetylcysteine and the ampicillin are compounded, the minimal inhibitory concentration of the ampicillin for main pathogenic bacteria, including escherichia coli, staphylococcus aureus, streptococcus agalactiae and streptococcus dysgalactiae, of the dairy cow mastitis can be remarkably reduced when being compared with that of single utilization of the ampicillin, so that the bacterium inhibition or bacterium killing effect of the ampicillin on the pathogenic bacteria is enhanced, and the pharmaceutical composition has a remarkable treatment effect on the dairy cow mastitis caused by the pathogenic bacteria; the pharmaceutical composition adopts a paste preparation and is injected into mammary chambers of dairy cows with the mastitis through a teat canal, so that the pathogenic bacteria can be rapidly killed; meanwhile, the pathogenic bacteria in the environment can be prevented from entering the mammary chambers and a prevention effect is realized; the pharmaceutical composition is easily absorbed and breast tissues of the dairy cows with the mastitis are not injured.

The invention relates to a primer combination capable of detecting three kinds of streptococcus simultaneously. The primer combination consists of upstream and downstream primers capable of detectingstreptococcus agalactiae, as shown in SEQ ID MO:1, 2, upstream and downstream primers capable of detecting streptococcus dysgalactiae, as shown in SEQ ID MO:3, 4, and upstsream and downstream primerscapable of detecting streptococcus iniae, as shown in SEQ ID MO:5, 6. A reagent kit consists of a DNA extraction reagent and a PCR reaction reagent, wherein the DNA extraction reagent consists of a TEbuffer solution, proteinase k, chloroform, isopropyl alcohol, ethanol and redistilled water, and the PCR reaction reagent contains a reaction buffer solution, dNTPs, DNA polymerase, the 6 primers anddeionized water. A method for detecting the three kinds of streptococcus simultaneously comprises the steps of extracting DNA in samples through the extraction reagent, performing PCR amplification on the DNA through the reaction reagent, and performing gel electrophoresis on amplification products. The three kinds of streptococcus can be detected simultaneously, detection is simple, quick, highin sensitivity and high in specificity, and the primer combination and the method can meet requirements for large-scale molecular epidemiology investigation and analysis of aquatic animals and qualitysafety detection of aquatic products.

The invention discloses a lincomycin hydrochloride oil solution and a preparation method and application thereof, and belongs to the field of medicine and pharmacy. The lincomycin hydrochloride oil solution overcomes the problem of the easy settlement of a mixed suspend lincomycin hydrochloride oil preparation. The lincomycin hydrochloride oil solution is simple in preparation process only needing common physical mixing without grinding process. The lincomycin hydrochloride oil solution is free of visible insolubles under a microscope, and can be sterilized through a 0.22 mum millipore filter film. The lincomycin hydrochloride oil solution product has good liquidity, and may not be hung on the wall, the appearance is single-phase, transparent and clear, clarity detection is acceptable, and after repeated freezing and thawing, preparation stratification and drug settlement phenomenon may not occur. The lincomycin hydrochloride oil solution has the advantages of simple process, stable quality and great application prospect. The lincomycin hydrochloride oil solution product is used for treatment of mastitis of lactating cows infected with staphylococcus aureus, streptococcus agalactiae, streptococcus dysgalactiae, streptococcus uberis escherichia coli and fungus and the like.

Disclosed are antibodies that block the binding of fibronectin protein to fibronectin. Also disclosed are site specifically-mutated and truncated peptide epitopes derived from the fnbA and fnbB genes of Staphylococcus aureus, the fnbA and fnbB genes of Streptococcus dysgalactiae, and the sfb gene of Streptococcus pyogenes, and nucleic acid segments encoding these peptides and epitopes. The anti-(fibronectin binding site) antibodies, peptides and epitopes that give rise to antibodies that block the binding of fibronectin binding proteins to fibronectin, and DNA segments encoding these proteins and are of use in various screening, diagnostic and therapeutic applications including active and passive immunization and methods for the prevention of streptococcal and staphylococcal colonization in animals or humans. These DNA segments and the peptides derived therefrom are proposed to be of use directly in the preparation of vaccines and also for use as carrier proteins in vaccine formulations.

The invention discloses a preparation method and application of a micro-capsule coated egg yolk antibody IgY resistant to main pathogenic bacteria of dairy cow mastitis. A compound mycoprotein antigen is prepared by selecting streptococcus agalactiae, streptococcus dysgalactiae, staphylococcus aureus and escherichia coli and is mixed with an equal quantity of freund's adjuvant to prepare a multivalence antigen immune complex, laying hens are immune by adopting repeatedly alternative immune procedures and a muscular and subcutaneous multipoint injection means, a specific yolk immunoglobulin IgY resistant to the main pathogenic bacteria of dairy cow mastitis, namely DP-BM-IgY, is separated and purified from collected egg yolk, then homogeneous emulsification is conducted on the specific yolk immunoglobulin and a sodium alginate water solution and an emulsifying agent soya bean salad oil containing span-80, the emulsified solution is dropwise added into an encystation solution prepared from CaCl2 and chitosan for coating and curing, and the coated micro-capsules can effectively protect the activity of the DP-BM-IgY and have enteric solubility. The preparation method utilizes a micro-capsule technology and selects the chitosan and sodium alginate as coating materials (natural polysaccharides), and the egg yolk antibody is internally taken and free of any toxic or side effect.

This invention discloses a lysin that can kill many species of Streptococci. A new lysin, ClyR, was constructed by the gene splicing method. The ClyR can effectively kill different species of Streptococci, including Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus suis, Streptococcus uberis, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus mutans, Streptococcus equi, and various Enterococci and Staphylococcus aureus. ClyR shows good stability and is not sensitive to EDTA and high concentration of NaCl. Moreover, the ClyR is active in a wide range of pH and maintains high activity in pH 5-11. Recombinant protein ClyR is well expressed in E. coli stain BL21 (DE3). High doses of ClyR showed no apparent toxicity in mice. Furthermore, administration of 0.8 mg per mouse once is able to completely protect the mouse infected with lethal doses of Group B Streptococci. The ClyR can be used alone or in combination with different forms of reagents and solutions, for the control of a variety of Streptococci and for the treatment of infections caused by these bacteria. It has a broad application prospect.

The present invention aims at the 16S-23S rRNA interval of the main pathogenic bacteria Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae and Escherichia coli of dairy cow mastitis, synthesizes 4 pairs of corresponding specific amplification primers, and applies a multiplex PCR reaction system to the cow udder detection of pathogenic bacteria. The lengths of the amplified fragments were 162bp for Streptococcus agalactiae, 264bp for Streptococcus dysgalactiae, 439bp for Staphylococcus aureus and 544bp for Escherichia coli. The multiplex PCR detection kit and detection method have strong specificity and sensitivity. It can detect and identify four kinds of pathogenic bacteria at the same time, so as to achieve the purpose of reducing the workload and cost of PCR diagnosis.

The invention provides a reagent kit for detecting streptococcus pyogenes. The reagent kit contains a guide RNA specifically targeted to a streptococcus pyogenes rpoB gene, an amplification primer pair, a hydration TwistAmp basic kit reaction drying ball, LbCas12a protein, a single-stranded DNA probe (ssDNA), a Ribonuclease Inhibitor and a buffer solution. When the reagent kit is used for detecting the streptococcus pyogenes, the detection specificity is high, and the streptococcus pyogenes can be distinguished from other streptococcus including streptococcus dysgalactiae and streptococcus agalactiae. Besides, when the RNA sequence is used for detection, time consumption is about 1 hour, a PCR instrument is not needed, requirements for equipment are low and are notably lower than those by a traditional bacteria culture method or a PCR sequencing method, and the clinical use value is large.

The GapC plasmin binding protein genes of Streptococcus dysgalactiae (S. dysgalactiae), Streptococcus agalactiae (S. agalactiae), Streptococcus uberis (S. uberis), Streptococcus parauberis (S. parauberis), and Streptococcus iniae (S. iniae) are described, as well as the recombinant production of the GapC proteins therefrom. Also described is the use of the GapC proteins from those species in vaccine compositions to prevent or treat bacterial infections in general, and mastitis in particular.

The GapC plasmin binding protein genes of Streptococcus dysgalactiae (S. dysgalactiae), Streptococcus agalactiae (S. agalactiae), Streptococcus uberis (S. uberis), Streptococcus parauberis (S. parauberis), and Streptococcus iniae (S. iniae) are described, as well as the recombinant production of the GapC proteins therefrom. Also described is the use of the GapC proteins from those species in vaccine compositions to prevent or treat bacterial infections in general, and mastitis in particular.

The GapC plasmin binding protein genes of Streptococcus dysgalactiae (S. dysgalactiae), Streptococcus agalactiae (S. agalactiae), Streptococcus uberis (S. uberis), Streptococcus parauberis (S. parauberis), and Streptococcus iniae (S. iniae) are described, as well as the recombinant production of the GapC proteins therefrom. Also described is the use of the GapC proteins from those species in vaccine compositions to prevent or treat bacterial infections in general, and mastitis in particular.

A method for hyaluronic acid having a low degradation rate in a subject body includes culturing Streptococcus dysgalactiae strain 9103 (KCTC11818BP) in a medium including a carbon source and a nitrogen source.

The invention provides a bacteriostasis method for selectively inhibiting gram-positive bacteria. The method uses small molecule PI-n as a bacteriostatic agent, the structure of the bacteriostatic agent is that biimidazole is connected with pyrene fluorophore through 1-4 methylene groups, and the bacteriostatic agent can selectively inhibit the growth of gram-positive bacteria, including staphylococcus aureus, staphylococcus saprophyticus, enterococcus faecalis, bacillus cereus, streptococcus dysgalactiae and the like, and particularly has a certain inhibition effect on clinical methicillin-resistant staphylococcus aureus (MRSA). The action mechanism is that positively charged PI-n molecules can be highly gathered on the surface of gram-positive bacteria, and under the action of a certain dosage of PI-n, the permeability of bacterial cell walls is changed, and bacteria die due to swelling and rupture, so that the bactericidal effect is achieved.

The invention provides a kit for detecting dairy cow mastitis pathogenic bacteria. Comprising a primer for amplifying staphylococcus aureus, a primer for amplifying staphylococcus epidermidis, a primer for amplifying streptococcus agalactiae, a primer for amplifying streptococcus dysgalactiae, a primer for amplifying streptococcus uberis, a primer for amplifying enterococcus, a primer for amplifying pseudomonas aeruginosa, a primer for amplifying enterobacter and a primer for amplifying escherichia coli. A primer for amplifying Klebsiella pneumoniae, a primer for amplifying listeria monocytogenes and a primer for amplifying mycoplasma bovis. The invention also provides a method for detecting cow mastitis by adopting the kit. The method has the advantages of being convenient to operate, free of special instruments and low in cost, not only can provide powerful help for accurate diagnosis and treatment of a farm, but also has the advantages of being high in detection efficiency and low in cost, is very suitable for basic-level use and has the condition of large-scale application and popularization.

The invention discloses a bactericidal preparation, which contains 0.0001-0.1 wt% of lysostaphin, 10-50 wt% of lysozyme, 1-15 wt% of a stabilizer, 0.1-0.5 wt% of a surfactant and the balance of water. The invention also discloses a bactericidal wet tissue, its preparation method and its application in treating and preventing milk cow mammary gland infection. The bactericidal wet tissue can be obtained by loading the bactericidal preparation on a carrier. The bactericidal wet tissue provided by the invention is used to kill various bacteria that cause milk cow mammary gland infection, and can achieve very significant effects, especially for E. , Corynebacterium pyogenes killing rate of more than 99%.

The invention discloses sturgeon source Streptococcus dysgalactiae and application thereof. The sturgeon source Streptococcus dysgalactiae is preserved at the China typical culture preservation centerof the Wuhan University (on a campus of the Wuhan University, No.299, Bayi Road, Wuchang District, Wuhan City, Hubei Province) in November 29th, 2018, the preservation number is CCTCC M 2018843, andthe classified name of a bacterial strain is Streptococcus dysgalactiae SDS. The invention further discloses the application of the sturgeon source Streptococcus dysgalactiae to a drug for preventionand control over sturgeon streptococcicosis. The drug can serve as a vaccine, the vaccine has good immune protection capability, sturgeon is subjected to immunization through the vaccine and thus canresist disoperation of streptococcosis of domestic main sturgeon breeding varieties, and market prospects are broad.

Provided is a Streptococcus dysgalactiae ID9103 strain having accession number KCTC11818BP, and a method of producing hyaluronic acid by culturing the strain to produce hyaluronic acid having an average molecular weight of 10,000,000 Da or more.

The recombinant production of Gap4, a chimeric GapC plasmin binding protein comprising the entire amino acid sequence of the Streptococcus dysgalactiae GapC protein in addition to unique amino acid sequences from the Streptococcus parauberis and Streptococcus agalactiae GapC proteins, is described. Also described is the use of Gap4 chimeric GapC protein in vaccine compositions to prevent or treat streptococcal infections in general and mastitis in particular.

The invention provides a detection gene chip and a detection method for the main pathogenic bacteria of cow mastitis. The gene chip includes a solid phase carrier and a specific detection probe fixed on the solid phase carrier, and the specific detection probe includes 6 kinds of universal primer sequences designed with the Escherichia coli 16S rDNA conserved region as the base sequence. A specific oligonucleotide hybridization probe designed in the variable region of the main pathogenic bacteria of dairy cow mastitis; the six main pathogenic bacteria of dairy cow mastitis are Streptococcus agalactiae, Streptococcus dysgalactiae, Escherichia coli, pneumoniae Klebsiella, Proteus mirabilis, and Staphylococcus aureus. The invention precisely limits the annealing temperature of the PCR reaction, optimizes the hybridization conditions of the PCR-amplified nucleic acid fragment and the gene chip, obtains a good hybridization effect, and finally obtains accurate detection results. The invention has good specificity, high accuracy rate, reliable stability, low false positive rate of detection results, simple operation steps and easy popularization and use.