17 results about "Brucella Vaccine" patented technology

The invention discloses an mRNA-LNPs Brucella vaccine, an expression vector and a preparation method thereof. The mRNA-LNPs Brucella vaccine comprises at least one of mRNA of an outer membrane protein BP26 and mRNA of an outer membrane protein Omp 25. According to the present invention, the antigen sequence is optimized, the prepared mRNA Brucella vaccine can stimulate the body to produce strong immunogenicity, and compared with the Brucella vaccine prepared before the sequence optimization, the antibody titer is significantly improved. A mouse immune experiment research result shows that high-titer IgG, IgG1 and IgG2a can be generated. Th1 type reaction, Th2 type reaction or mixed Th type reaction is successfully induced.

The application discloses a serological method for identifying Brucella A19-DeltaVirB12 vaccine immunized animals and infected animals, and belongs to the technical field of veterinary vaccine detection, comprising the step of identifying the antibodies of the serum by using the NH-AGID method. The method helps to perfect the animal Brucella A19-DeltaVirB12 vaccine prevention and control technical system, makes up for the short board that the A19-DeltaVirB12 vaccine immunized animals cannot be identified, and provides technical support for realizing the precise prevention and control of bovine brucellosis. The application of the method will accelerate the application of the Brucella A19-DeltaVirB12 vaccine to replace the traditional A19 vaccine, and opens a new milestone for the prevention and control of Brucella.

This invention provides a live attenuated Brucella vaccine strain M5Δ pyrE Its construction methods and applications belong to the field of biotechnology. The M5... pyrE The strain was obtained by knocking out Brucella mesenteriae strain M5. pyrE Genetically constructed and deposited at the China Center for Type Culture Collection. This invention discovers that... pyrE The gene is associated with Brucella virulence; knocking out this gene significantly weakens Brucella virulence and reduces its intracellular and mouse viability. Compared to the existing vaccine strain M5-9026, M5... pyrE The strain exhibits superior immunoprotective effects, effectively inducing humoral and Th1 cellular immune responses, and demonstrates good safety. The M5 strain constructed in this invention... pyrE This strain has promising potential as a candidate strain for a live attenuated vaccine to prevent brucellosis in animals.

This invention provides a primer-probe combination, reagents, methods, and applications for detecting Brucella vaccine strain RB51, belonging to the field of pathogen detection technology. The primer-probe combination for detecting Brucella vaccine strain RB51 of this invention includes a forward primer RB51-F, a reverse primer RB51-R, and a probe RB51-P; the forward primer RB51-F is shown in SEQ ID NO:1, the reverse primer RB51-R is shown in SEQ ID NO:2, and the probe RB51-P is shown in SEQ ID NO:3. The fluorescent PCR and digital PCR detection methods established based on the primer-probe combination of this invention have high sensitivity, good specificity, fast detection speed, and high throughput, enabling rapid detection of Brucella vaccine strain RB51, making differential diagnosis more accurate, economical, efficient, and convenient.

The present application provides a target point, a kit and a method for identifying Brucella vaccine strains and wild strains. The present application obtains a specific nucleotide sequence capable of specifically distinguishing Brucella vaccine strains and wild strains through a large number of research and analysis, which can be used as an effective target point for identifying Brucella vaccine strains and wild strains. The present application also provides a primer composition for multiplex RPA amplification of the target point, and a crRNA composition for detecting the specific nucleotide sequence based on the CRISPR / Cas12a technology. Using the crRNA composition for CRISPR / Cas12a detection of the product of the multiplex RPA amplification, the multiplex amplification and typing detection can be quickly and efficiently (about 35 min) completed, and the minimum detection limit is 5 copies / μL, which has good specificity and sensitivity.

The invention discloses an immortalized goat bone marrow-derived macrophage for expressing an SV40LT gene and application of the immortalized goat bone marrow-derived macrophage, and belongs to the technical field of bioengineering. The immortalized goat bone marrow-derived macrophage provided by the invention is obtained by knocking an immortalized gene SV40LT into a primary goat bone marrow-derived macrophage ROSA26 site through a CRISPR-Cas 9 system, and the immortalized goat macrophage provided by the invention is an immortalized goat macrophage cell strain which is suitable for being infected with salmonella and brucella and can be continuously and stably passaged; the method can be effectively applied to immunological research on goat brucella infection resistance and production of brucella vaccines.

The invention belongs to the technical field of biological medicines, and particularly relates to a Brucella recombinant multi-epitope vaccine and application thereof. The invention provides a recombinant multi-epitope vaccine protein of Brucella. The vaccine protein is a first vaccine variant rBMEV1 or a second vaccine variant rBMEV2. Results of embodiments prove that the vaccine provided by the invention can excite strong cross immune response aiming at main pathogenic species (such as sheep species, cattle species and pig species) of Brucella. Animal experiments prove that the antibody can induce a high-level and lasting specific antibody, can obviously activate response of CD4 + and CD8 + T cells and corresponding cell factors, and has good safety at the same time. According to the invention, the problems of narrow protection spectrum, dependence on exogenous adjuvants, single immune type and the like of the traditional brucella vaccine are solved, and a new generation of efficient, broad-spectrum and safe candidate vaccine strategy is provided for prevention and control of brucellosis.

The invention provides a target spot, a kit and a method for identifying a brucella vaccine strain and a wild strain. According to the invention, a specific nucleotide sequence capable of realizing specific region distribution of a brucella vaccine strain and a wild strain is obtained through a large amount of research and analysis, and can be used as a target spot for effectively identifying the brucella vaccine strain and the wild strain. The invention also provides a primer composition for carrying out multiple RPA amplification on the target spot, and a crRNA composition for detecting the specific nucleotide sequence on the basis of a CRISPR / Cas12a (Clustered Regularly Interspaced Short Palindromic Repeats / Cas12a) technology. The crRNA composition is used for carrying out CRISPR / Cas12a detection on the multiple RPA amplification product, multi-target amplification and typing detection can be rapidly and efficiently completed (about 35 min), the lowest detection limit is 5 copies / mu L, and the crRNA composition has good specificity and sensitivity.

The application belongs to the technical field of biotechnology, and provides a L7 / L12-PADRE sequence-multiple B cell epitope recombinant protein and kit for detecting anti-Brucella antibody. The recombinant protein is a multi-epitope tandem recombinant protein composed of Brucella ribosome L7 / L12 protein, PADRE polypeptide sequence and multiple B cell epitopes, and the amino acid sequence of the recombinant protein is shown as SEQ ID No. 1, and the antigenicity is good. The anti-Brucella antibody indirect ELISA (iELISA) detection method and kit established by taking the recombinant protein as a coating antigen have the characteristics of high sensitivity, strong specificity and good repeatability, are suitable for detecting the anti-Brucella antibody produced by the organism, can clearly determine the Brucella infection or Brucella vaccine immunization background, provide basic data for preventing and controlling the human-animal shared Brucella disease, and are simultaneously convenient for being applied to large-scale sample detection and epidemiological monitoring.

Provided are a system and method for preparing outer membrane vesicles with guaranteed immune effects. The system comprises a strain acquisition module, a strain screening module, an outer membrane vesicle yield promotion module, an outer membrane vesicle extraction module, an outer membrane vesicle purification module, an immune efficacy evaluation module, and a yield evaluation module, wherein the strain acquisition module is connected to the strain screening module, the outer membrane vesicle yield promotion module, and the outer membrane vesicle extraction module, respectively. By means of a genetic modification operation, the alr gene of Brucella is knocked out, which in turn improves the yield and immune efficacy of the outer membrane vesicles. Moreover, by means of strictly controlling the Brucella culture conditions, an inducing environment is established to induce Brucella to secrete outer membrane vesicles, thereby further improving the yield of outer membrane vesicles, and in turn obtaining outer membrane vesicles with a high yield and good immune effects, which meets the requirements of large-scale vaccine production, and provides a high-quality vaccine adjuvant for the research into, development of and application of Brucella vaccines.

The invention discloses a brucella vaccine as well as a preparation method and application thereof, and belongs to the field of biological medicines. According to the invention, Brucella P39 protein with an amino acid sequence as shown in SEQ ID NO.1 is used as an antigen, mRNA with a nucleotide sequence as shown in SEQ ID NO.2 is designed according to a coding gene of the P39 protein, and the Brucella mRNA vaccine is prepared. Experimental results show that the mRNA vaccine provided by the invention can efficiently induce humoral immunity and cellular immunity at the same time, and Brucella infection is prevented through multiple mechanisms. The mRNA vaccine disclosed by the invention is simple and convenient to prepare, uniform in immunogenicity, capable of inducing to generate consistent and effective immune response, and lasting in immune effect. The invention provides a new biological material for preventing and treating brucella in animal husbandry, and has outstanding practical value and wide application prospect.

The invention relates to detection of the cellular immune level of a Brucella vaccine, in particular to an antibody and a kit for detecting specific gamma interferon generated by immunizing cattle with the Brucella vaccine. The invention provides an antibody pair for detecting bovine gamma interferon through a double-antibody sandwich enzyme-linked immunosorbent assay. The antibody pair consists of a capture antibody and a detection antibody, the capture antibody is secreted by a hybridoma cell with the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.45838; the detection antibody is secreted by a hybridoma cell with the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.46325. The bovine gamma interferon ELISA detection method established based on the antibody pair has the advantages of high sensitivity, strong specificity and good repeatability, and can be used for detecting the production level of specific gamma interferon of Brucella vaccine immunized cattle so as to evaluate the cellular immune level of an organism.

The present application belongs to the field of microbial genetic engineering, and particularly relates to a brucellosis cellular immunity protein and application thereof. Firstly, a brucella protein with high immunogenicity is screened through antibody profiling technology; then, molecular biology technology is used to fuse and express the brucella protein gene sequence and the brucella T cell epitope and antigen gene sequence, so as to improve the ability of the antigen to induce immune response; then, the antigen is subjected to multistage purification after in-vitro expression, and the obtained purified protein is used as a stimulant; finally, the anticoagulated blood of an immunized animal is collected, the stimulant is added, and the mixture is incubated at 37 DEG C for 24 hours; the supernatant is collected by centrifugation, and the IL-17 level in the supernatant is detected by ELISA. The brucellosis cellular immunity protein is prepared into a reagent for detecting the IL-17 level, and the IL-17 is detected by ELISA, so that the protection level of an animal after brucella vaccine immunization can be effectively evaluated, the operation is simple and accurate, and the problem that the protection level of the vaccine after brucella vaccine immunization cannot be quickly evaluated is solved.

The invention provides a method for identifying a Brucella BA0711 vaccine strain and a wild strain, and in the method, a specific detection target gene for identifying the Brucella BA0711 vaccine strain is located in a 400 bp sequence as shown in SEQ ID NO. 7, and specific detection target genes for identifying other Brucella strains are located in a 606 bp sequence as shown in SEQ ID NO. 8. The invention also provides an application of the identification method, and a kit for detecting a vaccine strain and a wild strain of Brucella BA0711. The research aims at a Brucella BA0711 vaccine strain, and a specific molecular marker gene capable of regionally distributing the Brucella BA0711 vaccine strain and a wild strain is screened out. A specific primer and a probe are designed based on the marker gene. The method is a qPCR detection method capable of accurately identifying the brucella BA0711 vaccine strain, the brucella wild strain and mixed infection of the brucella BA0711 vaccine strain and the brucella wild strain. The method can solve the problem of lack of a rapid identification tool in production practice, and provides technical support for infection monitoring and precise purification of vaccine-immunized livestock groups.

The invention discloses an attenuated smooth Brucella strain and a preparation method and application of a vaccine, the attenuated smooth Brucella strain is obtained by deleting a bpe275 gene of a bovine species Brucella A19 strain, the strain is preserved in China General Microbiological Culture Collection Center (CGMCC) on July 15, 2025, the preservation unit address is CGMCC No.35193, No.1 yard, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC NO.35193. The classification name of the strain is A19 [delta] BPE275 bovine Brucella abortus, a specific fragment of 1431bp can be obtained, the smooth type characteristic is retained, and the sequence of the specific fragment is shown as SEQ ID No.11. The invention also discloses a preparation method of the Brucella abortus strain. A bpe275 gene deleted strain is constructed by adopting a Brucella vaccine A19 as a carrier, the survival ability is remarkably reduced, the toxicity is weakened in a mouse test, and the problem that the toxicity of a current Brucella vaccine is enhanced is solved; the smooth characteristic of the parent strain is maintained, the passage is stable, the bacterial form is not changed, and the organism can be stimulated to generate stronger cellular immunity.

The invention provides an enhanced brucella outer membrane vesicle (OMV) as well as a preparation method and application thereof. The core innovation of the preparation method is as follows: besides conventional culture conditions, culture conditions for simulating an intracellular living environment are additionally introduced to culture a 104M: Omp19 overexpression strain, and then the enhanced OMV which is more comprehensive in antigen composition and can better simulate a real infection state is extracted from the 104M: Omp19 overexpression strain. The preventive brucella vaccine prepared on the basis of the method does not need to add any adjuvant, and can generate a relatively good protection effect through two-needle immunization at a low dose (5 mu g).

The invention discloses a Brucella multi-epitope fusion protein, a vaccine, a preparation method and application, the Brucella multi-epitope fusion protein is designed based on a Brucella WbkC protein, a B cell epitope (LBL), a cytotoxic T cell epitope (CTL) and a helper T cell epitope (HTL) of the Brucella multi-epitope fusion protein are screened, and the Brucella multi-epitope fusion protein is prepared by connecting KK, AAY, GPGPG and GGGGS connecting peptides. A vaccine prepared from the fusion protein provided by the invention can provide better antigen-specific immune response than WbkC for immunizing mice. Therefore, the subunit fusion protein vaccine involved in the invention is expected to become a novel brucella vaccine, and has good application and development prospects.