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44 results about "Clostridium perfringens toxoid" patented technology

Clostridium perfringens alpha toxin is a toxin produced by the bacterium Clostridium perfringens (C. perfringens) and is responsible for gas gangrene and myonecrosis in infected tissues.

Clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method

The invention relates to a clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method. According to the clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method, alpha toxin protein obtained via prokaryotic expression is taken as an immunogen; monoclonal antibodies obtained via hybridoma technique are taken as detection antibodies and capture antibodies; reaction conditions are optimized via experiments; alpha toxin protein samples with a series of concentration are used for construction of a standard curve; the double-antibody sandwich ELIS method is established; and indexes of the double-antibody sandwich ELIS are verified. The clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method specifically comprises following steps: (1) prokaryotic expression of alpha toxin; (2) preparation of anti-alpha toxin monoclonal antibodies; (3) establishment of the double-antibody sandwich ELIS method; (4) establishment of the standard curve; and (5) performance evaluation on the double-antibody sandwich ELIS method. The clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method is excellent in specificity, and high in sensitivity and stability, is fast and convenient, and can be used for effective quantitative determination of alpha toxin in A-E type clostridium perfringens cultural supernatants; and the high-efficient detection method is provided for alpha toxin determination.
Owner:SHANDONG AGRICULTURAL UNIVERSITY

Monoclonal antibody blocking ELISA (enzyme-linked immuno sorbent assay) testing method of clostridium perfringens alpha toxin

The invention discloses a monoclonal antibody blocking ELISA testing method of a clostridium perfringens alpha toxin, which is used for early testing of the alpha toxin and provides an effective testing tool for early diagnosis, spreading prevention and subsequent research of clostridium perfringens. The method comprises the following two steps: I, establishing the monoclonal antibody blocking ELISA testing method; and II, determining a result judgment criterion. The monoclonal antibody blocking ELISA testing method which has good specificity and stability and high sensitivity, is quick, simple and convenient, and tests the alpha toxin efficiently is established for the first time, and the method is simple to implement and high in testing speed and can achieve quantitative testing and massive testing. In a word, monoclonal antibody blocking ELISA is expected to play an important role in prevention and treatment of clostridium perfringens diseases.
Owner:NORTHEAST AGRICULTURAL UNIVERSITY

Bovine type-A clostridium perfringens subunit vaccine and preparation method and application thereof

ActiveCN107596361AStrong immune responseOvercoming the problem of incomplete inactivationAntibacterial agentsBacterial antigen ingredientsAntigenAlpha-toxin
The invention provides a bovine type-A clostridium perfringens subunit vaccine and a preparation method and an application thereof. The subunit vaccine comprises a bovine type-A clostridium perfringens alpha-toxin protein truncation body and a pharmacologically acceptable adjuvant. The bovine type-A clostridium perfringens alpha-toxin protein truncation body is an antigen determinant amino acid sequence from 255th to 372nd amino acids of alpha-toxin protein. The subunit vaccine has the following advantages that 1) safety problems due to uncompleted inactivation are not existed; 2) the qualityis controllable, and the difference between the batches is not existed; 3) the production equipment and space requirement is low, the expression level is high, and the cost is low; and 4) toxin dispersion risk is not existed, and the safety of operators can be guaranteed.
Owner:NOVO BIOTECH CORP

Clostridium perfringens alpha, beta 1, beta 2 and epsilon coexpression vector and construction method and expression method thereof

The invention discloses a clostridium perfringens alpha, beta 1, beta 2 and epsilon toxin coexpression vector; vectors are respectively pETDuet-1 and pRSFDuet-1 expressed in the same host bacteria, and the vectors are respectively connected with clostridium perfringens alpha and epsilon toxin encoding genes and clostridium perfringens beta 1 and beta 2 toxin encoding genes. On the basis, the invention also discloses a construction method and an expression method of the coexpression vector.
Owner:LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI

Toxin attenuation mutant for epsilon toxin of clostridium perfringens and application of toxin attenuation mutant

ActiveCN104560780AGood immunogenicity and immunoprotectionAchieve complete attenuationAntibacterial agentsBacterial antigen ingredientsAromatic amino acidsMutant
The invention discloses a toxin attenuation mutant for epsilon toxin of clostridium perfringens and an application of the toxin attenuation mutant for the epsilon toxin of the clostridium perfringens. An epsilon toxin mutant for attenuating toxin in cells or animals is obtained by mutating tyrosine at a site 71 of mature toxin of the epsilon toxin of wide clostridium perfringens into non-aromatic amino acids. The invention further discloses a recombinant expression vector and a recombinant host cell which contain coding genes of the toxin attenuation mutant for the epsilon toxin of the clostridium perfringens. According to the recombined and expressed toxin attenuation mutant for the epsilon toxin of the clostridium perfringens, the complete toxin attenuation of mice in vitro and in vivo can be achieved, and good immunogenicity and immunizing protection are presented in a mouse model. The toxin attenuation mutant for the epsilon toxin of the clostridium perfringens and the coding genes of the toxin attenuation mutant can be used for preparing subunit vaccines of the epsilon toxin of the clostridium perfringens or subunit vaccines of multivalent clostridium toxin.
Owner:HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI

Genetically engineered vaccine of epsilon toxin of clostridium perfringens and application thereof

The invention discloses a genetically engineered vaccine of epsilon toxin of clostridium perfringens and an application thereof. The genetically engineered vaccine of epsilon toxin of clostridium perfringens is obtained by the following steps: firstly, designing a specific primer; carrying out PCR (Polymerase Chain Reaction) amplification to obtain an epsilon gene segment, and cloning to a pMD18-T Vector; selecting a positive recombinant, and guiding a recombinant plasmid into an acceptor strain to successfully construct a recombinant strain BL21 (DE3)-epsilon; and finally, emulsifying the prepared epsilon-toxin protein and an aluminum hydroxide adjuvant in a volume ratio of 9:1, and adding 0.01% of thiomersalate to obtain the genetically engineered vaccine of epsilon toxin of clostridium perfringens. The genetically engineered vaccine of epsilon toxin of clostridium perfringens can be put into industrial production easily, simple to operate, has good safety and the like, and can be applied to preventing fetal intestinal diseases of animal such as lambs, sheep, goats, calves and chinchilla, caused by D type clostridium perfringens.
Owner:SHANDONG AGRICULTURAL UNIVERSITY

Humanized single-chain antibody 8B of clostridium perfringens alpha-toxin

The invention discloses a humanized single-chain antibody 8B of clostridium perfringens alpha-toxin. The humanized single-chain antibody 8B is screened from a phage antibody library Source bioscience, is a full-humanized antibody of the clostridium perfringens alpha-toxin and can be used for achieving a toxin neutralization treating effect, overcoming various side effects of a heterologous antibody and eliminating complex steps and high cost of humanization modification on the heterologous antibody; and the single-chain antibody is small in molecular weight, strong in in-vivo penetrability and capable of rapidly reaching to damaged tissues and cells to exert an antitoxin effect, so that the double aims of economy and high efficiency are achieved. The alpha-toxin has a certain homology with other types of toxins of clostridium perfringens, so that the antibody drug possibly generates inhibiting and treating effects for other types of toxins and can also be used as a detecting reagent to detect the clostridium perfringens alpha-toxin.
Owner:MILITARY VETERINARY RES INST PLA MILITARY MEDICAL ACAD OF SCI

Clostridium perfringen alpha toxin genetic engineering vaccine and application thereof

The invention relates to a clostridium perfringen alpha toxin genetic engineering vaccine and application thereof. The clostridium perfringen alpha toxin genetic engineering vaccine is prepared by designing a specific primer, implementing polymerase chain reaction (PCR) amplification on A-type clostridium perfringen DNA to obtain an alpha toxin gene segment, cloning the gene segment onto a pMD18-T Vector, selecting a positive recon and linking the positive recon onto a PET-28 alpha (+) plasmid; introducing the recombinant plasmid to a receptor strain to successfully construct a recombinant strain BL21 (DE3)-alpha; emulsifying the prepared alpha toxin protein and an aluminum hydroxide adjuvant at a volume ratio of 9:1, and adding 0.01% thiomersal to obtain the clostridium perfringen alpha toxin genetic engineering vaccine. The clostridium perfringen alpha toxin genetic engineering vaccine is easy for realization of industrial production, simple to operate and good in safety, and is capable of effectively preventing such diseases as animal necrotic enteritis, enterotoxemia, and human and animal traumatic gas gangrene caused by the A-type clostridium perfringens.
Owner:SHANDONG AGRICULTURAL UNIVERSITY

Clostridium perfringens type B exotoxin, preparation method thereof, toxigenic culture medium and application of clostridium perfringens type B exotoxin

ActiveCN109554420AToxic performance is stableStabilize Toxic EffectsAntibacterial agentsBacterial antigen ingredientsVaccine ProductionBacillus perfringens
The invention discloses a clostridium perfringens type B exotoxin, a preparation method thereof, a toxigenic culture medium and an application thereof. Each 1000 ml of the toxigenic culture medium isprepared from the following raw materials by weight: 15-25 g of peptone, 3-5 g of yeast extract, 3-4 g of NaCl, 5-10 g of Na2HPO4.12H2O, 5-10 g of dextrin or 5-10 g of coarse dextrin, and the balanceof water for injection. The virulence of the clostridium perfringens toxin liquid produced by the method according to the present invention is up to 1 mouse MLD of no more than 0.0006 mL, and the virulence is 3.3 times than the vaccine production standard. Moreover, a triple inactivated vaccine against bradsot, struck (or lamb dysentery) and enterotoxemia prepared by the clostridium perfringens type B exotoxin has serum neutralizing titer in rabbits and sheep equal to or higher than the relevant standard.
Owner:JINYUBAOLING BIO PHARM CO LTD

Clostridium perfringen alpha toxin genetic engineering vaccine and preparation method thereof

The invention provides a clostridium perfringen alpha toxin genetic engineering vaccine and a preparation method thereof. Compared with wild type alpha toxins, clostridium perfringen alpha toxin recombination protein is characterized in that the 176th site histidine of an amino acid sequence is mutated to obtain asparagine. The clostridium perfringen alpha toxin genetic engineering vaccine is prepared from detoxifcation clostridium perfringen alpha toxin recombination protein of self-induction secretory expression. The vaccine has the advantages of being safer, better in immune efficacy, simpler in technology, lower in cost and the like, and can effectively solve the problems that in the prior art, the production technology of the vaccine is complexer, and the cost is higher.
Owner:JIANGSU ACADEMY OF AGRICULTURAL SCIENCES

Porcine epidemic diarrhea-porcine clostridial enteritis bivalent subunit oral vaccine and preparation method thereof

The invention belongs to the technical field of biology and provides a porcine epidemic diarrhea-porcine clostridial enteritis bivalent subunit oral vaccine and a preparation method thereof. The porcine epidemic diarrhea-porcine clostridial enteritis bivalent subunit oral vaccine comprises two antigens, a vaccine adjuvant and an oral adjuvant, wherein the two antigens are an inactivated porcine epidemic diarrhea virus S protein and an inactivated porcine clostridium perfringens beta toxin protein respectively. With the vaccine adopted, a pig can simultaneously obtain the porcine epidemic diarrhea virus antibody and the porcine clostridium perfringens beta toxin antibody, so that the pig can be prevented from being infected with epidemic diarrhea and clostridial enteritis. The vaccine is taken orally, so that an immunization route is changed; the immunological stress of the pig can be reduced; the side effect of the vaccine is slight; and therefore, the immune effect of the vaccine is effectively improved. The preparation method of the oral vaccine provided by the invention is simple and easy to operate and does not need special production equipment. The production cost of the oralvaccine is lower.
Owner:天康生物制药有限公司

Humanized single-chain antibody 7D of clostridium perfringens alpha-toxin

The invention discloses a humanized single-chain antibody 7D of clostridium perfringens alpha-toxin. The humanized single-chain antibody 7D is screened from a phage antibody library Source bioscience, is a full-humanized antibody of the clostridium perfringens alpha-toxin and can be used for achieving a toxin neutralization treating effect, overcoming various side effects of a heterologous antibody and eliminating complex steps and high cost of humanization modification on the heterologous antibody; and the single-chain antibody is small in molecular weight, strong in in-vivo penetrability and capable of rapidly reaching to damaged tissues and cells to exert an antitoxin effect, so that the double aims of economy and high efficiency are achieved. The alpha-toxin has a certain homology with other types of toxins of clostridium perfringens, so that the antibody drug possibly generates inhibiting and treating effects for other types of toxins and can also be used as a detecting reagent to detect the clostridium perfringens alpha-toxin.
Owner:MILITARY VETERINARY RES INST PLA MILITARY MEDICAL ACAD OF SCI

Establishment and application of surface display C type clostridium perfringens alpha and beta toxin protein recombinant lactobacillus plantarum

The invention relates to surface display C type clostridium perfringens alpha and beta toxin protein recombinant lactobacillus plantarum. The C type clostridium perfringens alpha toxin gene sequence,beta1 toxin gene sequence and beta2 toxin gene sequence are inserted in a genome of lactobacillus plantarum. The invention further relates to an establishment method and application of the recombinantlactobacillus plantarum. Through a PCR method, alpha, beta1 and beta2 toxin genes are successfully cloned on the whole genome of C type clostridium perfringens (CVCC 1158), a corresponding recombinant plasmid is established and electrically transferred into lactobacillus plantarum NC8, three strains of recombinant lactobacillus plantarum are obtained after induction expression, the oral mode is selected for immunizing a rat, the immune effect of recombinant bacteria is evaluated, the result shows that when the rat is immunized by orally taking the recombinant lactobacillus plantarum, the ratbody can be stimulated to generate humoral immunity and can also be stimulated to generate cellular immunity, and a good protection effect on an animal body is realized.
Owner:HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY

Indirect ELISA method of clostridium perfringens beta2 toxin antibody

The invention discloses an indirect ELISA (enzyme-linked immuno sorbent assay) detection method for a clostridium perfringens beta 2 toxin antibody. The indirect ELISA detection method comprises the following steps: step 1, establishing an indirect ELISA method for the clostridium perfringens beta 2 toxin antibody; and step 2, determining a result judgment standard. The invention belongs to the technical field of pathogenic microorganism detection, and particularly provides an indirect ELISA system for detecting a clostridium perfringens beta2 toxin antibody, which is good in specificity, high in sensitivity and good in repeatability, and provides an effective detection means for the clostridium perfringens infection condition of animals. The indirect ELISA detection method for the clostridium perfringens beta2 toxin antibody has a wide application prospect in the aspects of clinical serum epidemiological investigation and vaccine immune effect evaluation, and can be used for rapidly detecting the clostridium perfringens beta2 toxin antibody.
Owner:NINGXIA UNIVERSITY

Capture ELISA (enzyme-linked immunosorbent assay) detection method for Clostridium perfringens Alpha-toxin antibody

The invention relates to the field of animal bacteriology and provides a capture ELISA (enzyme-linked immunosorbent assay) detection method for a Clostridium perfringens Alpha-toxin antibody; in the method, prepared type-A Clostridium perfringens exotoxin is used as an antigen that is used for immunizing a rabbit to obtain rabbit serum and for acting as a binding antigen for capture ELISA after being concentrated and purified; the detection method comprises the steps of (1) enveloping; (2) sealing; (3) adding an antigen; (4) adding a sample to be tested; (5) adding a secondary antibody; (6) developing color; (7) ending; (8) judging results. The detection method has the advantages of high specificity, high sensitivity, good repeatability, low cost and the like; in addition, the detection method is suitable for detecting samples in large batch, and an effective and simple serological diagnostic method is provided for the detection and research on such toxin.
Owner:SHANDONG AGRICULTURAL UNIVERSITY

Preparation method and application of veterinary-use A type clostridium perfringens toxins

The invention discloses a preparation method and application of veterinary-use A type clostridium perfringens toxoids. The preparation method of the toxoids includes the following steps that A type clostridium perfringens production strains are inoculated into a culture medium for fermentation and culture, a fermentation product is obtained, then the fermentation product is inactivated and detoxified in L-lysine and a formaldehyde aqueous solution under the pH of 6.8, and centrifugal supernatant of the successfully inactivated and detoxified qualified bacterial solution refer to toxoids. The culture medium includes casein peptone, yeast extract powder, CaCl2, ZnSO4 7H2O, Na2HPO4 12H2O, KH2PO4, and glucose. The toxicity of the A type clostridium perfringens toxoids prepared by the method can be raised to be 12.5 times of the standard for seedling cultivation, the neutralization titer, in first and second immune serum of domestic rabbits, of the toxoids can be raised to 4 and 50 times ofa traditional process respectively and maximally, and the ratio of the serum titer to cost can be raised to 48.8 and 1216.4 times of the traditional process.
Owner:CHINA INST OF VETERINARY DRUG CONTROL

Clostridium perfringens epsilon toxin attenuated mutant and its application

The invention discloses a toxin attenuation mutant for epsilon toxin of clostridium perfringens and an application of the toxin attenuation mutant for the epsilon toxin of the clostridium perfringens. An epsilon toxin mutant for attenuating toxin in cells or animals is obtained by mutating tyrosine at a site 71 of mature toxin of the epsilon toxin of wide clostridium perfringens into non-aromatic amino acids. The invention further discloses a recombinant expression vector and a recombinant host cell which contain coding genes of the toxin attenuation mutant for the epsilon toxin of the clostridium perfringens. According to the recombined and expressed toxin attenuation mutant for the epsilon toxin of the clostridium perfringens, the complete toxin attenuation of mice in vitro and in vivo can be achieved, and good immunogenicity and immunizing protection are presented in a mouse model. The toxin attenuation mutant for the epsilon toxin of the clostridium perfringens and the coding genes of the toxin attenuation mutant can be used for preparing subunit vaccines of the epsilon toxin of the clostridium perfringens or subunit vaccines of multivalent clostridium toxin.
Owner:HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI

Poultry vaccine for clostridium perfringens

The present disclosure relates to nanoparticle compositions for use as vaccines against Clostridium perfringens in poultry which causes necrotic enteritis in poultry. Such compositions include one or more Clostridium perfringens extracellular proteins entrapped in a polyanhydride or chitosan nanoparticle. The one or more Clostridium perfringens extracellular proteins may include one or more Clostridium perfringens toxins, such as, for example, alpha toxin (CPA), beta toxin (CPB), epsilon toxin (ETX), iota toxin (ITX), perfringolysin O (PFO), enterotoxin (CPE), beta2 toxin (CPB2), or NetB toxin. In some aspects, the composition further includes a Salmonella enteritidis flagellar protein. The present invention also includes methods for the oral delivery of one or more Clostridium perfringens extracellular proteins to the mucosal membrane of the intestinal tract of a bird of the order Galliformes.
Owner:UNIV OF GEORGIA RES FOUND INC +1

Clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method

The invention relates to a clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method. According to the clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method, alpha toxin protein obtained via prokaryotic expression is taken as an immunogen; monoclonal antibodies obtained via hybridoma technique are taken as detection antibodies and capture antibodies; reaction conditions are optimized via experiments; alpha toxin protein samples with a series of concentration are used for construction of a standard curve; the double-antibody sandwich ELIS method is established; and indexes of the double-antibody sandwich ELIS are verified. The clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method specifically comprises following steps: (1) prokaryotic expression of alpha toxin; (2) preparation of anti-alpha toxin monoclonal antibodies; (3) establishment of the double-antibody sandwich ELIS method; (4) establishment of the standard curve; and (5) performance evaluation on the double-antibody sandwich ELIS method. The clostridium perfringens alpha toxin double-antibody sandwich ELIS quantitative determination method is excellent in specificity, and high in sensitivity and stability, is fast and convenient, and can be used for effective quantitative determination of alpha toxin in A-E type clostridium perfringens cultural supernatants; and the high-efficient detection method is provided for alpha toxin determination.
Owner:SHANDONG AGRICULTURAL UNIVERSITY

Clostridium perfringens beta toxin mutant protein as well as preparation method, application and vaccine thereof

ActiveCN111944028ANon-toxicSuccessfully attenuatedAntibacterial agentsBacterial antigen ingredientsMutated proteinClostridium perfringens beta toxin
The invention provides clostridium perfringens beta toxin mutant protein as well as a preparation method, application and a vaccine thereof, and relates to the field of biological products. The clostridium perfringens beta toxin mutant protein is prepared by mutating 212th arginine and 266th tyrosine of wild clostridium perfringens beta toxin with an amino acid sequence as shown in SEQ ID NO.3 into alanine, and the toxicity of the clostridium perfringens beta toxin mutant protein is remarkably reduced compared with that of the wild clostridium perfringens beta toxin. Meanwhile, good immunogenicity is also reserved.
Owner:天康制药股份有限公司

Clostridium perfringens epsilon toxin mutant protein as well as preparation method, application and vaccine thereof

The invention provides a clostridium perfringens epsilon toxin mutant protein as well as a preparation method, application and a vaccine thereof, and relates to the technical field of biology. The clostridium perfringens epsilon toxin mutant protein includes alanine mutated by 151-position amino acid of clostridium perfringens epsilon toxin shown in an amino acid sequence SEQ ID NO.3. Compared with a wild type, the clostridium perfringens epsilon toxin protein has significantly reduced toxicity and good immunogenicity. The vaccine can alleviate technical problems such as complex production process, high cost, complex antigen, inter-batch difference and large side effects of inactivated vaccine.
Owner:天康制药股份有限公司

A Recombinant Fusion Protein of Avirulent Tetanus Toxin and Clostridium perfringens β Toxin

The invention relates to non-toxic tetanus toxin and clostridium perfringens beta toxin recombined fusion protein. By means of codon optimization, clostridium tetani fragments C and the recombined fusion protein containing multiple clostridium perfringens beta toxin mutants with amino acid mutation, the immunogenicity of two kinds of toxin protein is reserved to the maximum extent, and potential biosafety hazards caused by single amino acid mutation are avoided. The recombined fusion protein can be used for preparing unit vaccines of clostridium tetani, B type clostridium perfringens and C type clostridium perfringens; compared with a current commercial clostridium natural toxin inactivated vaccine in China, the recombined fusion protein has the advantages of being simple in preparation process, low in immunizing dose, excellent in vaccine potency and the like, the biosafety risk in the vaccine production process are greatly reduced, and the recombined fusion protein is an ideal candidate vaccine antigen for upgrading and updating the three kinds of clostridium toxin vaccines. When the recombined fusion protein is used for preparing a combined vaccine with other antigens, the combined vaccine can be prepared without increasing the use dose of the combined vaccine.
Owner:CHINA INST OF VETERINARY DRUG CONTROL

Clostridium perfringens alpha toxin visual detection method

The invention discloses a Clostridium perfringens alpha toxin visual detection method. A smartphone image processing technology and silicon dioxide microsphere antibody coupling are combined to detectfood-borne clostridium perfringens alpha toxin (CPA). According to a double-antibody sandwich principle, the prepared CPA polyclonal antibody is coupled with silicon dioxide microspheres, CPA fluorescence detection is performed through antibody-labeled FITC fluorescence, and finally, the CPA content is detected in real time in combination with an image processing technology of a smartphone APP. Compared with a traditional clostridial toxin detection method, the nano-microsphere combined intelligent mobile phone CPA detection system is a more sensitive, stable and convenient detection method,and a new method is provided for detection of natural CPA in food.
Owner:TIANJIN UNIV
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