104 results about "Streptococcus mastitidis" patented technology

The invention provides proteins from group B streptococcus (Streptococcus agalactiae) and group A streptococcus (Streptococcus pyogenes), including amino acid sequences and the corresponding nucleotide sequences. Data are given to show that the proteins are useful antigens for vaccines, immunogenic compositions, and / or diagnostics. The proteins are also targets for antibiotics.

The invention provides proteins from group B streptococcus (Streptococcus agalactiae) and group A streptococcus (Streptococcus pyogenes), including amino acid sequences and the corresponding nucleotide sequences. Data are given to show that the proteins are useful antigens for vaccines, immunogenic compositions, and / or diagnostics. The proteins are also targets for antibiotics.

The invention relates to an aquaculture animal disease control technology and in particular relates to a tilapia mossambica source streptococcus agalactiae low virulent strain and application thereof. The tilapia mossambica source streptococcus agalactiae low virulent strain is YM001 and is preserved in CCTCC (China Center For Type Culture Collection) with the preservation number of CCTCCM2014045. The tilapia mossambica source streptococcus agalactiae low virulent strain is subjected to activation culture and fermentation culture, bacterial cells of a 106-109CFU / mL low virulent strain YM001 and a sterile phosphate buffered solution are mixed as a low virulent live vaccine used for preventing and controlling a tilapia mossambica streptococcus disease, the injection medication concentration for each tilapia mossambica is 105-108CFU, the oral medication concentration for each tilapia mossambica is 108-109CFU, and the soaking medication concentration is 107-108CFU / mL. The invention relates to strain identification, toxicity test, toxicity return safety and stability, immunogenicity and immune protection effect of a low virulent strain, and the tilapia mossambica streptococcus disease vaccine prepared by adopting the low virulent strain is good in immune protection effect and strong in safety and stability.

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 discloses a preparation method and application of a tilapia mossambica-derived streptococcus agalactiae recombinant CBP protein vaccine. The recombinant CBP protein vaccine is prepared from tilapia mossambica-derived streptococcus agalactiae recombinant CBP protein. The amino acid sequence of the tilapia mossambica-derived streptococcus agalactiae recombinant CBP protein is shown as a sequence 2. Tilapia mossambica-derived streptococcus agalactiae is used as a research object and is cloned into a Cbp gene from a tilapia mossambica-derived streptococcus agalactiae genome; the tilapia mossambica-derived streptococcus agalactiae is connected with a pET28a(+) carrier to construct an expression vector, and prokaryotic expression and purification are carried out in E.coliBL21(DE3), so as to obtain the recombinant CBP protein vaccine with high specificity and immune protective efficacy of 56.35 percent+ / -8.09 percent; a foundation is laid for immune prevention and treatment of tilapia mossambica streptococcus agalactiae diseases.

The invention discloses a method for constructing an anti-mammitis transgenic mouse model and a special vector for the method. An anti-mammitis-related protein composition comprises lysostaphin and peptidoglycan endolysin B30; the peptidoglycan endolysin B30 derives from a Streptococcus agalactiae phage B30, and an amino acid sequence of the peptidoglycan endolysin B30 is sequence 2 in a sequence table; and an amino acid sequence of the lysostaphin is sequence 3 in the sequence table. The experiment proves that a transgenic plasmid for mammary gland-specific expression of the lysostaphin and the peptidoglycan endolysin B30 is constructed by using a mammary gland-specific expression vector; and the anti-mammitis transgenic mouse model is constructed by a microinjection method, a transgenic mouse with an anti-bovine mastitis gene is obtained, and a technical model is provided for further producing anti-mammitis transgenic calves.

The present invention relates to polypeptides, epitopes and antibodies directed to these epitopes, more particularly to the Sip polypeptide of Group B streptococcus (GBS), also called Streptococcus Agalactiae which may be used to prevent, diagnose and / or treat streptococcal infection.

The invention discloses new pleuromutilin derivatives which have a structural formula (I) or a structural formula (II) shown in the specification, wherein in the formula (I), when n=0, R1 is Cl, CH3, OCH3 or NH2; and when n=1, R1=H, and R2=NH2. The compounds have favorable inhibiting action on Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and Streptococcus mastitidis; the antibacterial activity of one compound having a phenyl group with ortho-substituting groups or para-substituting groups (such as Cl, CH3, OCH3 or NH2) is superior to the antibacterial activity of one compound having a phenyl group with meta-substituting groups; and part of compounds having para-substituting aryl groups or ortho-substituting aryl groups are the same with valnemulin in the antibacterial activity on Staphylococcus epidermidis or Streptococcus mastitidis, and can be used for the preparation of antibacterial drugs. The synthesis method of the compounds has the advantages that the raw materials are accessible, the price is low, the operation process is simple, the products are easy to separate and purify, the yield is high and the total yield is 35-45%.

The invention discloses a preparation method of a pleuromutilin compound, a polymorphism and a preparation method of the polymorphism. The pleuromutilin compound comprises a 14-O-[(4-amino-6-hydroxy-pyrimidine-2-yl) thio acetyl] matrix shown in a structural formula (i) and / or a 14-O-[(4-amino-6-one-pyrimidine-2-yl) thio acetyl] matrix shown in a structural formula (ii). The novel pleuromutilin compound disclosed by the invention has obvious inhibiting effects on multiple drug resistant bacteria such as methicillin resistant staphylococcus aureus and staphylococcus epidermidis as well as common pathogenic bacteria such as escherichia coli and streptococcus mastitidis in veterinary clinics, so that the pleuromutilin compound can be applied to preparation of antibacterial drugs and particularly anti-drug resistant bacterium drugs.

The invention relates to bacillus cereus NY5 capable of effectively inhibiting tilapia-source streptococcus agalactiae. The strain is preserved in the China General Microbiological Culture Collection Center (CGMCC) and the preservation number is CGMCC No.9372. The bacillus cereus is aerobic bacteria and can effectively inhibit growth of tilapia-source streptococcus agalactiae. Through a nitrate nitrogen experiment, removal efficiency can reach 100%. The strain also has functions of casein hydrolysate and starch and has passed safety detection. In a water body with concentration of bacillus cereus NY5 being 2*107cfu / ml and under the condition of injecting 100 microliters of a NY5 bacterium solution at the concentration of 2.1*106cfu / ml, no death or other abnormal phenomena happen to tilapia. According to the invention, a useful strain source is provided for effectively inhibiting growth of tilapia-source streptococcus agalactiae and efficiently degrading nitrite nitrogen, protolysate and starch under the aerobic condition, and the application range of bacillus cereus in the aspect of functions is widened. The bacillus cereus NY5 has high application value.

The invention discloses a preparation method and application of a tilapia source streptococcus agalactiae recombinant GroEL protein vaccine. The recombinant GroEL protein vaccine is prepared by tilapia source streptococcus agalactiae recombinant GroEL protein. An amino acid sequence of the tilapia source streptococcus agalactiae recombinant GroEL protein is as shown in SEQ ID NO.2. In the invention, tilapia source streptococcus agalactiae is taken as a research object, a GroEL gene is obtained from a tilapia source streptococcus agalactiae genome by cloning, the cloned GroEL gene is connected with a pET28a(+) carrier to construct an expression vector, and the vector is subjected to prokaryotic expression and purification in E.coli BL21(DE3) to obtain the recombinant GroEL protein vaccine with strong specificity and an immune protective efficacy of 68.61+ / -7.39 percent, so that a foundation is laid for immune prevention and treatment of a tilapia streptococcus agalactiae disease.

The invention discloses a detection primer set of Streptococcus agalactiae. The detection primer set of Streptococcus agalactiae comprises a primer pair hylB, a primer pair ponA and a primer pair cfb, wherein the primer pair hylB comprises a primer hylB-F of which the nucleotide sequence is as shown in SEQ ID NO.1 and a primer hylB-R of which the nucleotide sequence is as shown in SEQ ID NO.2; the primer pair ponA comprises a primer ponA-F of which the nucleotide sequence is as shown in SEQ ID NO.3 and a primer ponA-R of which the nucleotide sequence is as shown in SEQ ID NO.4; the primer pair cfb comprises a primer cfb-F of which the nucleotide sequence is shown in SEQ ID NO.5 and a primer cfb-R of which the nucleotide sequence is shown in SEQ ID NO.6. The invention further discloses a detection kit containing the primer set and a multiplex PCR detection method using the detection kit. The primer set and the detection method disclosed by the invention are good in specificity, high in sensitivity, simple and fast, high-efficient and accurate, suitable for rapid inspection and quarantine of Streptococcus agalactiae in pollution-free aquatic products, and can be directly applied to early monitoring and early warning of aquaculture diseases. The lowest concentration of Streptococcus agalactiae DNA detected by the detection kit is 7.74*10<-3>ng / uL, and the detection kit can achieve minimally invasive sampling without bacterial culture, and requiring small quantity of samples for detection.

This invention relates to new vaccines against microorganisms based on antigenically mimetic peptides. The invention also relates to methods of discovering such mimetic peptides by first screening peptide-display phage libraries with antibodies against the microbial carbohydrates(s) of interest to locate antigenically mimetic peptides. Vaccines against Group B Streptococcus, or Streptococcus Agalactiae, are preferably produced using this method.

The invention discloses a tilapia mossambica streptococcus agalactiae oral attenuated freeze-dried vaccine and its preparation method. According to the invention, tilapia mossambica streptococcus agalactiae attenuated bacterial strain YM001 is performed with steps of enlarge cultivation, thalline collection and freeze-drying preparation to obtain the oral attenuated vaccine. The immunizing dose, immunization program and a vaccine usage method of the vaccine can be determined through tests. The immune efficacy, security and maneuverability of the vaccine can be detected and confirmed through field tests, the result shows that the vaccine has the advantages of high protection effectiveness, good security, convenient usage and low cost, the vaccine can be used for preventing tilapia mossambica streptococcus agalactiae, and has high business value.

The invention discloses recombinant lactococcus lactis based on the fbsA gene of Streptococcus agalactiae. The recombinant lactococcus lactis comprises the fbsA gene of Streptococcus agalactiae or segments of the fbsA gene, the fbsA gene or the segments of the fbsA gene preferably has (or have) a sequence shown in SEQ ID No. 1, the deposit number of the recombinant lactococcus lactis is CCTCC M 2020140, and expression of recombinant FbsA protein with a theoretical molecular weight of 34.6 kDa can be induced under the culture condition that niusin exists. The invention also provides a vaccine for the Streptococcus agalactiae disease of Oreochromis sp., and the vaccine is prepared from the recombinant lactococcus lactis, and is capable of producing protective antibodies in Oreochromis sp. after Oreochromis sp. is fed with the vaccine in an immune mode of oral gavage, so that Oreochromis sp. is assisted to resist infection of Streptococcus agalactiae effectively.

The invention discloses a rapid separation and identification kit for streptococcus agalactiae. In the kit, the following reagents are included: sheep defibering blood plate, 3% H2O2, staphylococcus aureus strains, a specific PCR (polymerase chain reaction) solution, and positive control DNA (deoxyribonucleic acid). The rapid separation and identification kit for streptococcus agalactiae provided by the invention has the following beneficial effects that bacterial morphological observation, catalase test, CAMP (cyclic adenylic acid) reaction and PCR detection are used together, so that identification procedures can be simplified, the specificity is strong, and the sensitivity is increased by more than 15%; and moreover, the identification cost is low, the time consumption is short (only needing 3 days), and streptococcus agalactiae strains can be obtained by separation and accurately identified. The kit can be used for separation and identification for the samples of the milk samples of human or dairy cows, cervical mucus, vaginal swabs and the like.

The present invention relates to polypeptides, epitopes and antibodies directed to these epitopes, more particularly to the Sip polypeptide of Group B Streptococcus (GBS), also called Streptococcus agalactiae which may be used to prevent, diagnose and / or treat streptococcal infection.

The invention provides a LAMP (Loop-Mediated Isothermal Amplification) primer group, a micro-fluidic chip and a kit for detecting reproductive tract pathogenic microorganisms, and belongs to the technical field of reproductive tract infection detection. The LAMP primer group comprises a streptococcus agalactiae detection primer group, an enterococcus faecalis primer group, a gardnerella vaginosisprimer group, a Candida albicans primer group and a Chlamydia trachomatis primer group. The above LAMP primer group is fixed on the micro-fluidic chip. The LAMP primer group, the micro-fluidic chip and the kit can quickly, sensitively and accurately detect streptococcus agalactiae, enterococcus faecalis, gardnerella vaginosis, Candida albicans and Chlamydia trachomatis by high repeatability.

The invention discloses a kit for directly extracting DNA of three pathogenic bacteria from milk. The kit mainly comprises a solution A, wherein the solution A contains EDTA-2Na, NaOH, SDS and NaCl. The kit is simple in reagent component; lysozyme and proteinase k are not used; the high-toxicity organic solvents, such as phenol, chloroform and ethyl alcohol, are not used; the cost is low and the operation is simple and convenient. Based on the well-designed extracting solution and a microwave method, the invention also discloses a method for directly extracting DNA of three pathogenic bacteria from milk. The three pathogenic bacteria comprise escherichia coli, staphylococcus aureus and streptococcus agalactiae. According to the method, the cell splitting effect is improved, the conventional extracting steps are simplified, only one EP tube is required during the milk processing process, the whole extracting process is shortened within 30min, an acquired template can be directly used for PCR reaction, the PCR amplification product is detected through agarose gel electrophoresis, and then the detection of the sample is finished within short time.

The invention discloses a method for constructing a high-yield glutathione recombinant strain and application, and belongs to the field of bioengineering. The method for constructing the high-yield glutathione recombinant strain is characterized in that corynebacterium glutamicum for synthesizing a large amount of glutamic acid is used as an original strain, and a gene gshF from streptococcus agalactiae or genes gshA and gshB from escherichia coli are introduced to endow glutathione with the synthesis capability; further genome is subjected to deletion of genes aceD, mcbR and sdaA; a truncatedgene serA, a gene pgk, genes cysE and cysK derived from the escherichia coli and a gene cysE derived from arabidopsis thaliana are expressed at the same time, and a cysteine pathway is strengthened;and as the corynebacterium glutamicum can synthesize a large amount of the glutamic acid and the cysteine pathway is strengthened, the glutathione can be highly produced only by adding glycine in a fermentation process of obtained engineering bacteria.

The invention relates to a culture-independent semiquantitative detection kit of tilapia streptococcus agalactiae, which consists of a reagent A, a reagent B, a reagent C, a reagent D, a reagent E, a product specification and a test result record card. The kit can be used for directly detecting whether blood of tilapia carries the streptococcus agalactiae without pure culture by a method provided by the invention, and semiquantitatively detect the quantity of the streptococcus agalactiae in the blood.

The invention discloses streptococcus agalactiae BibA recombinant protein, as well as a coding gene, a preparation method and an application thereof. The streptococcus agalactiae BibA recombinant protein provided by the invention is a protein sequence as shown in sequence No. 2. The invention also provides the coding gene, the preparation method and application of the protein. The streptococcus agalactiae BibA recombinant protein has the beneficial effects that streptococcus agalactiae is used as a research object, streptococcus agalactiae DNA is cloned to obtain a BibA gene, the cloning and sequencing of the gene sequence are completed, the prokaryotic expression and immunizing protection of the streptococcus agalactiae are researched, a vaccine development candidate antigen with high specificity and immune protection force is obtained, and the foundation is laid for prevention of streptococcus agalactiae diseases. The streptococcus agalactiae BibA recombinant protein has the advantages that the streptococcus agalactiae BibA gene is obtained by cloning, BibA recombinant expression plasmids are constructed, the BibA recombinant protein is obtained, westernblot detection results show that the BibA recombinant protein has relatively good reactogenicity, and animal (rat) experiments prove that the recombinant protein has certain protectiveness.

The invention relates to an aquaculture animal vaccine, in particular to an attenuated live vaccine prepared from a tilapia mossambica streptococcus agalactiae natural attenuated strain TFJ0901. The streptococcus agalactiae natural attenuated strain TFJ0901 is preserved in China Center for Type Culture Collection of Wuhan University, China, and the preservation number is CCTCC NO: M 2016792. The ingredients of the attenuated live vaccine are live cells of the natural attenuated strain TFJ0901 and a phosphate buffer solution, and the cell concentration is 10<6>-10<8> CFU / ml. The vaccine has high safety and immunogenicity and can effectively prevent tilapia mossambica streptococcus agalactiae.

The invention relates to recombinant Lactococcus lactis comprising a fusion gene of a sip gene and a pgk gene of Streptococcus agalactiae, and the fusion gene is introduced into the Lactococcus lactisby a plasmid vector. The fusion gene has a DNA sequence as shown in SEQ No. 1. The plasmid vector is a pNZ8148 vector. The lactococcus lactis is Lactococcus. lactis NZ9000. The invention further provides a tilapia mossambica streptococcus agalactiae disease vaccine. The tilapia mossambica streptococcus agalactiae disease vaccine comprises the recombinant lactococcus lactis. Wherein the recombinant lactococcus lactis is the recombinant lactococcus lactis induced by nisin. The vaccine is the vaccine administrated by oral administration and intragastric administration. The tilapia mossambica streptococcus agalactiae disease vaccine comprises the recombinant lactococcus lactis with the concentration of 2*10 <10> cfu mL <-1>, and the administration dosage is 200 [mu]L.

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.

The invention belongs to the technical field of aquaculture preparations, and discloses a feed additive for preventing streptococcus agalactiae, being prepared from the following components by mass percent: 10-15 percent of coptis chinensis, 7-15 percent of honeysuckle, 7-15 percent of isatis root, 10-20 percent of Houttuynia cordata, 20-40 percent of black soya bean and 0-20 percent of dendrobe. The additive is added into tilapia feed, has a highest immune protective rate of 95.21 percent for tilapia, and has practical and wide application values; the survival rate of the tilapia is 93.67 percent.

The invention belongs to the technical field of biology and specifically discloses the application of galectin-8 derived from Siniperca chuatsi in preparation of a bacteriostatic agent. The amino acidsequence of the galectin-8 is shown as SEQ ID NO. 1. It is found that the Siniperca chuatsi galectin-8 (ScGal8) has important antibacterial activity on typical aquatic animal pathogens; and an antibacterial experiment shows that the galectin-8 has no antibacterial activity on Aeromonas salmonicida (A. Salmonicida) and Edwardsiella tarda (E. Tarda), but has a relatively obvious inhibition effect on Streptococcus agalactiae (S. agalactiae) and Flavobacterium cloumnare (F. cloumnare), so the fish galectin-8 (rScGal8) has good application prospects in the treatment of gill rot diseases of aquaticanimals caused by Flavobacterium cloumnare.

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 method for screening candidate bacterial strains from fish streptococcus agalactiae vaccines. The method comprises the steps: separation and breed conservation of tilapia streptococcicosis agalactiae epidemic strains, identification of the epidemic strains and the establishing of a pathogenic library, PFGE genotype analysis of epidemic strains, toxicity determination of PFGE genotype representative strains, and immunogenicity and protection domain test of the PFGE genotype representative strains so as to obtain a candidate bacterial strains combination of a tilapia streptococcicosis agalactiae immunoprophylaxis vaccine in China. The technology has the characterizes of strong pertinence, high screening efficiency, remarkable effect and the like, and the screen candidate bacterial strains combination of the tilapia streptococcosis agalactiae immunoprophylaxis vaccine in China can protect 90% of genotypes and 96.47% of epidemic strains in the pathogenic library. The technology provides a new method for screening the candidate bacterial strains from the fish streptococcus agalactiae vaccines, is suitable for screening the candidate bacterial strains from the fish streptococcus agalactiae vaccines, and has significance and using value in immune prevention and control on fish streptococcicosis.

The invention relates to a tilapia streptococcus agalactiae vaccine and a preparation method thereof. According to bioinformatics principles, a part of polypeptide sequences of a surface anchoring protein of streptococcus agalactiae are analyzed and screened as an antigen for preparing the vaccine, in addition, a carbon nanotube carried surface protein system is constructed by using a chemical synthesis technology, and the selected polypeptide antigen is in nano combination with functionalized single-wall carbon nanotubes, so that a tilapia streptococcus agalactiae subunit vaccine is prepared.The polypeptide antigen screened by the invention is capable of remarkably increasing the antibody level of inoculated tilapia when being compared with an inactivated vaccine, and is capable of resisting attack of streptococcus agalactiae of high toxicity. In addition, the polypeptide antigen screened by the invention can be tightly and uniformly connected with surfaces of functionalization modified carbon nanotubes, and compared with independent polypeptide antigen groups, the polypeptide antigen is capable of remarkably improving the immunity of the tilapia.