20 results about "Hog cholera virus" patented technology

The invention relates to the technical field of biology, and aims to provide a monoclonal antibody 7A12 for identifying gene type 2 swine fever as well as a preparation method and an application method of the monoclonal antibody 7A12. The monoclonal antibody comprises Ig structural domains VH CDR1, VH CDR2 and VH CDR3 of a heavy chain of the antibody, and Ig structural domains VL CDR1, VL CDR2 and VL CDR3 of a light chain of the antibody, wherein the amino acid sequences of the VH CDR1, the VH CDR2 and the VH CDR3 are respectively as shown in SEQ ID NO: 1 to SEQ ID NO: 3; the amino acid sequences of the VL CDR1, the VL CDR2 and the VL CDR3 are respectively as shown in SEQ ID NO: 4 to SEQ ID NO: 6. The antibody provided by the invention has the advantages of high titer, strong specificity, high affinity and the like; the kit has the advantages of high sensitivity and strong specificity, has no cross reaction with the type 1 swine fever vaccine strain, and can identify and diagnose swine fever gene type 2 wild strain infection and vaccine immunity in serology; the method can be further used for establishing a blocking ELISA kit for identifying the gene type 2 hog cholera, and can effectively diagnose and purify the gene type 2 hog cholera virus.

The invention discloses a swine fever and porcine parvovirus bivalent subunit vaccine and a preparation method thereof. The vaccine comprises a first recombinant protein encoded by a first gene, a second recombinant protein encoded by a second gene and a pharmaceutically acceptable carrier. The first gene has a sequence as shown in SEQ ID NO: 1 or an increased or reduced sequence thereof. And the second gene has a sequence as shown in SEQ ID NO: 2 or an increased or reduced sequence thereof. An antigen E2 protein of a hog cholera virus (CSFV) and a VP2 protein of a porcine parvovirus (PPV) are taken as double targets, a recombinant SC-E2 protein with a SpyCatcher tag and a recombinant ST-VP2 protein with a SpyTag tag are respectively expressed in insect cells through a recombinant baculovirus vector, double-antigen covalent assembly is realized in vitro, and the constructed bivalent subunit vaccine can be used for simultaneously preventing and controlling two epidemic diseases and has a good application prospect. And the vaccine has the advantages of high safety, strong immunogenicity, high prevention and control efficiency, easiness in large-scale production and the like.

The invention discloses a hog cholera virus detection system based on cloud analysis, which comprises an image acquisition unit, a data processing cloud, a disease monitoring unit and an early warning management unit, and is characterized in that a live pig limb image is acquired through the image acquisition unit, and an image analysis model is constructed through the data processing cloud to analyze the live pig limb image; the hog cholera risk suspected degree is comprehensively evaluated from two perspectives of vascular risk and tissue edema, so that auxiliary diagnosis is carried out on hog cholera virus detection; the disease monitoring unit is used for monitoring the hog cholera virus for a long time, the deterioration or recovery condition of the disease condition of the sick pig is analyzed, a sick pig health management scheme is generated, and the targeted treatment effect of the sick pig is improved; and the early warning management unit globally analyzes the virus risk and generates a live pig breeding management scheme, so that corresponding overall management and control are performed for the hog cholera virus infection degree of the live pig breeding base, the early warning management efficiency of the live pig breeding base is improved, and the live pig breeding loss and the management cost are reduced.

The invention relates to molecular biology and biomedicine technologies, and aims to provide a preparation method and application of an efficient targeting swine fever nano vaccine. The invention provides an APC cell targeted hog cholera virus self-assembled protein nanoparticle, which is obtained by carrying out fusion connection expression on a recombinant plasmid of an APC cell targeted nanoparticle mi3-PTC with a sequence as shown in SEQ ID NO.3 and a hog cholera virus antigen with a sequence as shown in SEQ ID NO.5-12, and further mixing with pharmaceutically acceptable auxiliary materials to prepare a vaccine product for preventing and / or treating hog cholera virus infection. The APC cell targeting polypeptide can be displayed on the surface of the nanoparticle mi3, and the nanoparticle is directionally carried to the APC cell expressing CD163, so that the internalization, storage and distribution capabilities of the nanoparticle are remarkably improved in vivo and in vitro, and the vaccine can remarkably improve the immune response to the swine fever antigen.

The invention discloses a hog cholera subunit vaccine and a preparation method thereof, and belongs to the technical field of hog cholera subunit vaccines.The hog cholera subunit vaccine is a pure-line hog cholera lapinized virus strain CSFV CC-1 which is highly adaptive to rabbit source cells or common cells, single in genotype and stable in inheritance and is obtained through a hog cholera lapinized virus strain through a virus dilution technology. The swine fever subunit vaccine is developed by matching the treated and extracted virus subunit antigen stock solution with the subunit antigen adjuvant, so that the possibilities of virus dispersion, homologous recombination and variation can be effectively reduced; the immunogenicity of antigens is enhanced, the onset time of immunoreaction is shortened, pigs can be effectively prevented and protected from being infected by hog cholera viruses, the infection rate and breeding loss are greatly reduced, infected animals and immune animals can be effectively distinguished, and the elimination of potential hog cholera viruses in pig farms is effectively accelerated.

The invention relates to the technical field of prevention and control of epidemic diseases of direct and indirect contact or aerosol transmission of African swine fever and the like, in particular to an epidemic prevention type pig outgoing system which comprises a pig outgoing area and a pig receiving area, the pig outgoing area and the pig receiving area are isolated through a partition piece, a closed pig driving channel is arranged between the pig outgoing area and the pig receiving area, and the closed pig driving channel is communicated with the partition piece. The pig inlet end of the closed pig driving channel is located in the pig outlet area, the pig outlet end of the closed pig driving channel is located in the pig receiving area, and an air filtering system is arranged in the closed pig driving channel. The problem that the transmission of the hog cholera virus cannot be effectively prevented and controlled in the process of leaving the pigs can be solved.

The invention provides a hog cholera virus E2 fusion protein and a preparation method and application of a vaccine. Compared with the prior art, the E2 fusion protein provided by the invention has the beneficial effects that the C terminal of the E2 protein is replaced by the fusion protein of a porcine epidemic diarrhea virus transmembrane region, the blocking rate of the fusion protein to the hog cholera virus is relatively high, and polynucleotide synthesized by depending on the fusion protein can efficiently express the fusion protein; the mRNA vaccine prepared on the basis of the polynucleotide has a good immune protection effect on pigs, is not influenced by the existence of maternal antibodies, is high in safety and has a wide application prospect.

The invention belongs to the technical field of animal pathogen molecular biology, and particularly relates to a dual quantitative fluorescent primer probe composition, a kit and an identification method for identifying hog cholera virus. Aiming at the current popular strains covering 3 genotypes, 11 gene subtypes and the like of the hog cholera gene, the invention selects 5UTR and NS3 gene sequences of hog cholera virus to design and synthesize specific primers and probes aiming at the two fragments; the specific primer has no non-specific amplification on African swine fever virus, pseudorabies virus, porcine reproductive and respiratory syndrome virus, porcine circovirus type II, porcine epidemic diarrhea and bovine viral diarrhea. The invention establishes a method for identifying the classical swine fever virus wild strain and the vaccine strain and develops a kit convenient for diagnosis, and the kit and the identification method can simultaneously identify the classical swine fever virus wild strain and the vaccine strain, have the characteristics of high sensitivity, strong specificity, good repeatability and good stability, and have extremely strong application prospects.

The utility model discloses a culture device capable of continuously harvesting hog cholera virus liquid, which relates to the technical field of hog cholera virus liquid culture and comprises a base for improving the stability of the device, one side of the base is fixedly connected with a lead for supplying power to the base, and a mounting shell is inserted into the base. The bottom of an inner cavity of the mounting shell is fixedly connected with a driving motor, and the top of a rotating shaft of the driving motor is fixedly connected with a hog cholera virus liquid culture inner tank. A nutrient solution and hog cholera viruses are injected into the hog cholera virus liquid culture inner tank along the liquid adding pipe for virus culture, and when hog cholera virus liquid is collected, the control panel can control the output power of the driving motor, so that the output motor drives the inner tank to rotate at a high speed, and the hog cholera viruses are cultured in the hog cholera virus liquid culture inner tank. The classical swine fever virus liquid in the classical swine fever virus culture inner tank is centrifugally separated, and the settled classical swine fever virus liquid is filtered through the microporous filter element, so that the classical swine fever virus liquid culture device has an impurity removal effect and a virus liquid purification function.

The invention discloses a primer probe combination and a kit for fluorescent quantitative PCR (Polymerase Chain Reaction) detection of African swine fever virus and application of the primer probe combination and the kit. The primer probe combination is a specific primer probe combination 1 or a specific primer probe combination 2. The nucleotide sequence of the specific primer probe combination 1 is as shown in SEQ ID NO.1-3, and the nucleotide sequence of the specific primer probe combination 2 is as shown in SEQ ID NO.4-6. The qPCR detection method with strong specificity and high sensitivity is established, the lower detection limit can reach 0.87 copies / L, the sensitivity of the qPCR detection method is equivalent to that of a WOAH recommendation method, and the detection rate of weak positive samples is higher; the kit has no cross reaction with common swine pathogens such as hog cholera virus and pseudorabies virus, and has good specificity. The method provides reliable technical support for early diagnosis and precise prevention and control of African swine fever.

The invention discloses a swine fever, porcine reproductive and respiratory syndrome and African swine fever virus antibody fluorescence joint detection test paper card, a preparation method and a use method, a PVC backing (9) is installed on a card shell (10), and a nitrocellulose membrane (7) is laid on the PVC backing (9); a third detection line (5) coated with hog cholera virus E2a protein, a second detection line (4) coated with porcine reproductive and respiratory syndrome virus N1 protein, a first detection line (3) coated with African hog cholera virus P30 protein and a quality control line (6) of an anti-his tag protein antibody are sequentially marked on the nitrocellulose membrane (9). The kit has the advantages that the time-resolved immunofluorescent microspheres are used as tracers, the antigens of the three pig diseases are marked at the same time at a time, and meanwhile, the detection sensitivity and the detection efficiency are remarkably improved.

The invention discloses a mammalian cell strain, a method for preparing a hog cholera virus soluble E2 recombinant antigen by using the mammalian cell strain and application of the mammalian cell strain. The mammalian cell expression system with different cell generations can stably express a large amount of soluble CSFV-E2 recombinant protein, so that the production cost for producing the hog cholera E2 protein is effectively reduced, and further the mammalian cell expression system can be applied to production of the hog cholera E2 protein subunit vaccine.

The invention relates to the technical field of molecular biological detection, and particularly discloses a kit capable of simultaneously detecting eight porcine respiratory pathogens and application. The invention specifically relates to detection of hog cholera virus, porcine reproductive and respiratory syndrome virus, pseudorabies virus, circovirus type 2, African hog cholera virus, swine influenza A virus, streptococcus suis and swine pasteurella multocida. According to the combined detection composition provided by the invention, a dual quadruple fluorescent PCR analysis method is utilized, and different pathogens are detected by detecting target spots on the different pathogens, so that target genes of eight pathogens can be rapidly detected at a time, and detection and differential diagnosis are more efficient, economical, simple and convenient; the kit has the advantages of high sensitivity, good specificity, rapidness, high flux, simplicity and convenience in operation and the like.

The invention discloses a classical swine fever virus E2 protein labeled subunit vaccine and a serological differential diagnosis kit matched with the classical swine fever virus E2 protein labeled subunit vaccine. The invention belongs to the field of medical preparations, and relates to a classical swine fever virus E2 protein labeled subunit vaccine and a serological differential diagnosis kit matched with the classical swine fever virus E2 protein labeled subunit vaccine. Wherein the hog cholera virus recombinant E2 protein is obtained by mutating the epitope of the E2 protein of a hog cholera virus vaccine strain LPC, and the amino acid sequence of the mutant protein is SEQ ID No: 1 or a fusion protein obtained by connecting a protein tag to the N terminal or / and C terminal of the SEQ ID No: 1. The E2 marked subunit vaccine prepared by using the recombinant E2 protein has good immunogenicity, the diagnostic kit is prepared by using the E2 protein of the classical swine fever virus vaccine strain LPC and the monoclonal antibody of the DIDA55 recognition epitope of the classical swine fever virus E2 protein, and serological identification of vaccination of pigs without the marked vaccine or wild virus infection is realized, so that purification of classical swine fever is promoted.

The invention provides a construction method of a suspension HEK293 cell line capable of stably expressing classical swine fever virus vaccine strain E2 protein. The invention constructs a suspension HEK293 cell line for expressing hog cholera virus vaccine strain E2 protein. A cell stable passage test proves that when an HEK-E2 cell line can be stably passed for 10 generations, the expression quantity of the E2 protein in a culture supernatant is stable, the yield is stably kept at 1.0-1.2 mg / ml, the purity of the E2 protein obtained by affinity chromatography purification reaches 95%, double-antibody sandwich ELISA determines that the E2 protein has good reactogenicity, a 206 adjuvant emulsified E2 protein immune pig can induce a high-level serum antibody, and the E2 protein can be used for immunizing the HEK-E2 cell line for immunizing the HEK-E2 cell line for immunizing the HEK-E2 cell line for immunizing the HEK-E2 cell line. And after the virulent virus is attacked, complete protection can be achieved. The HEK-293 cell line constructed by the invention can stably secrete and express the CSFV E2 protein, and has an application prospect of being developed into a recombinant E2 genetic engineering vaccine.

The invention relates to the technical field of biology, in particular to a hog cholera virus E2 protein mutant and application thereof. The hog cholera virus E2 protein mutant comprises the following amino acid mutation sequence (1) or (2): (1) the hog cholera virus E2 protein is obtained by mutating the 15th site and the 25th site of the hog cholera virus E2 protein amino acid sequence; (2) mutating the 172nd site and the 323rd site of the amino acid sequence of the hog cholera virus E2 protein; wherein the amino acid sequence of the hog cholera virus E2 protein is as shown in SEQ ID NO. 1. The expression quantity of the hog cholera virus E2 protein mutant disclosed by the invention can reach 117-134 mu g / mL; the immune effect of the E2 protein mutant is better than that of the original sequence, and the positive rate of the E2 protein mutant after mice are immunized for 28 days reaches 100%.

The invention discloses a CHO cell culture method for expressing hog cholera virus E2 fusion protein. The CHO cell culture method comprises the following steps: subculture; and carrying out gradient cooling type fed-batch culture. The inoculation day is the 0th day, 3% of the material is supplemented on the 3rd day, 3% of the material is supplemented on the 4th day, 4% of the material is supplemented on the 5th day, 3% of the material is supplemented on the 6th day, 3% of the material is supplemented on the 7th day, 3% of the material is supplemented on the 8th day, 3% of the material is supplemented on the 10th day, and 3% of the material is supplemented on the 12th day; culturing in a gradient cooling manner, when the initial culture temperature is 36.5-37.5 DEG C and the cell density is 10.0-18.0 * 10 < 6 > cells / mL, cooling to 34-35 DEG C, cooling to 32-33 DEG C after 24 hours, cooling to 30-31 DEG C after 48 hours, maintaining until the 14th day or the cell viability is lower than 70%, harvesting supernatant, and finishing the culture. According to the culture process disclosed by the invention, the cell culture density and motility rate are improved, and the expression quantity of the E2 protein is remarkably improved, so that the production efficiency is improved, and the production cost is reduced.

The application discloses a CaO@SiO2 / multi-walled carbon nanotube composite material, a preparation method and application, and the preparation method comprises the following steps: step 1: CaO nanoparticles are dispersed in a solvent, a silicon source is added, and the CaO@SiO2 nanocomposite material is obtained through sufficient stirring reaction; and step 2: the CaO@SiO2 nanocomposite material and the multi-walled carbon nanotube are dispersed in a solvent, and the colloidal solution is obtained through ultrasonic mixing, and the colloidal solution is the required CaO@SiO2 / multi-walled carbon nanotube composite material; the electrochemical sensor prepared from the composite material obtained by the application can specifically detect hog cholera virus, the binding capacity of the composite material and a virus specific recognition element is improved, the interference of impurities such as impurities in a sample matrix is reduced, and the detection performance is excellent.

The invention belongs to the field of biology, and particularly relates to a PK-15 suspension cell line capable of efficiently breeding porcine circovirus and hog cholera virus, the PK-15 suspension cell line is proposed to be named as a porcine kidney cell suspension cell line in taxonomy, is preserved in Guangdong Microbial Culture Collection Center, has a preservation number of GDMCC NO.66284, is preserved on May 19, 2025, and has a preservation number of CGMCC NO.66284. The PK-15 suspension cell line can be used for efficiently breeding porcine circovirus and hog cholera virus. The preservation address is the fifth floor of building 59, No.100 courtyard, Xianlie Middle Road, Guangzhou City, Guangdong Province. The cell line is obtained through domestication by a suspension method originally created by the applicant, is named as PK15-WS04, and is preserved in Guangdong Microbial Culture Collection Center. After the PK-15 suspension cell line is subjected to 50 generations of passage, cells of each generation grow aseptically and are free of exogenous virus pollution, and the sensitivity of the PK-15 suspension cell line to porcine circovirus and hog cholera virus is not reduced; the immunogenicity of the virus prepared by each generation of cells is stable; the cell line can be used as a reserve cell line for vaccine production. Meanwhile, the invention also discloses a related application of the PK-15 suspension cell line.

The invention provides a method for preparing a hog cholera virus specific polyclonal antibody, and belongs to the technical field of veterinary biological products. The preparation method comprises the following steps: performing centrifugation, concentration, dilution, ultra-high-speed centrifugation and density gradient centrifugation on hog cholera virus to obtain non-hog cholera virus with relatively high purity, then combining virus liquid with a rabbit anti-hog cholera virus polyclonal antibody to obtain a complex, performing centrifugation, collecting supernate, adding the supernate into a serum-coated ELISA plate, and performing incubation at 37 DEG C for 1 hour, the detection result shows that the concentration of the specific polyclonal antibody reaches 1.0 mg / ml or above, and the specific polyclonal antibody does not react with other proteins in serum after being diluted by 1000 times or above, so that the specific polyclonal antibody shows relatively high specificity and is beneficial to accurate diagnosis and research.