127 results about "Pseudorabies" patented technology

The invention relates to the technical field of biology, in particular to a pseudorabies virus gD recombinant protein, a subunit vaccine and a preparation method and application thereof.The gD recombinant protein sequentially comprises a signal peptide, a connecting peptide, the 21st-350th site of the gD protein, an enzyme cutting site and a His tag from the N end to the C end, the amino acid sequence of the 21st-350th site of the gD protein is shown as SEQ ID No.4, and the amino acid sequence of the 21st-350th site of the gD protein is shown as SEQ ID No.6. The pseudorabies virus gD recombinant protein has high immunogenicity, the prepared subunit vaccine can generate a specific antibody with neutralizing activity, mice do not die after challenge, the protection rate is 100%, the protection rate after challenge of pigs is 100%, pathological sections only show slight pathological changes, and the pseudorabies virus gD recombinant protein has wide application prospects.

The invention discloses a preparation method and application of porcine pseudorabies virus multi-epitope polypeptide, and belongs to the technical field of veterinary biological products. The multi-epitope polypeptide comprises a cytotoxic T cell epitope, a helper T cell epitope and a linear B cell epitope; the cell epitopes are screened from key immunogenic proteins of the pseudorabies virus by using an immunoinformatics method. The porcine pseudorabies virus multi-epitope polypeptide designed by the invention is high in safety, has better antigenicity and physicochemical properties, can be efficiently expressed, and can be recognized by a pseudorabies virus specific antibody. By applying the porcine pseudorabies virus multi-epitope polypeptide, high-level specific antibodies and cellular immune response can be continuously induced to generate, strong protective efficacy is generated, and mice can be protected from death when infected with a lethal dose of porcine pseudorabies virus. Therefore, the multi-epitope polypeptide disclosed by the invention can be used as an alternative antigen of a porcine pseudorabies virus vaccine, the use of active virus is avoided, and a tool is provided for eradication of porcine pseudorabies virus.

The invention discloses application of naringenin in preparation of a medicine for preventing and treating diseases caused by pseudorabies virus, and belongs to the technical field of biological medicine. The naringenin has good anti-PRV activity in vivo and in vitro. The naringenin can inhibit virus proliferation of the pseudorabies virus in an intracellular replication stage, reduce expression of PRV gB protein, lower expression of TLR4 and P65 protein to inhibit activation of an NF-kB pathway and lower expression of an inflammatory gene, and reduce nucleation of the NF-kB / P65 protein to achieve anti-inflammatory and antiviral effects. In-vivo experiments prove that naringenin can delay the disease time of mice, reduce the virus load in organs of the mice infected by PRV and relieve pathological changes of tissues and organs of the mice, and in addition, the survival rate of the mice infected by viruses is improved by adjusting the serum cell factor level of the mice. The naringenin provides a new strategy for preventing and treating PRV infection.

The invention belongs to the technical field of pseudorabies virus prevention and control, and discloses application of BAY87-2243 in preparation of a drug for inhibiting pseudorabies virus proliferation, and the application is as follows: firstly, PK-15 is infected with a PRV-GD-WH strain, total RNA of cells is taken, and cDNA is obtained through reverse transcription; the change of the transcriptional level of a glycolysis key gene in a cell is detected through fluorescent quantitative PCR (Polymerase Chain Reaction) of an SYBR Green method. BAY87 is further used for acting on the HIF-1alpha, the regulation and control effect of the HIF-1alpha on glycolysis is verified, and finally, the virus load in cells is detected through SYBR Green fluorescent quantitative PCR (Polymerase Chain Reaction) and the inhibition effect of BAY87-2243 on PRV proliferation is determined through TCID50. Meanwhile, the invention also provides a pseudorabies virus inhibition drug.

The present application relates to the field of veterinary diagnostic technology, and specifically to a probe composition, a primer composition, and a triple fluorescence PCR kit and related methods for differential diagnosis of three genes of different strains of African swine fever composed of the above-mentioned probe composition and primer composition. The kit includes primer probes specific to the B646L gene, the CD2v gene, and the I177L gene. The present application designs fluorescent PCR primers and probes for the B646L gene, the CD2v gene, and the I177L gene, respectively, and establishes a triple fluorescence PCR detection method, which can quickly identify African swine fever virus strains and gene-deficient strains. At the same time, it has no cross-reaction with swine fever virus, porcine reproductive and respiratory syndrome virus, pseudorabies virus, porcine epidemic diarrhea virus, porcine circovirus, porcine parvovirus, healthy whole blood, oropharyngeal swabs, nasal swabs, anal swabs, environmental swabs, pig tissues, etc., and has good specificity.

The invention discloses preparation and application of pseudorabies virus circular RNA (Ribonucleic Acid). The circular RNA comprises a coding element for coding any one of gD protein, gB protein, gC protein and gK protein of the pseudorabies virus; preferably, the coding element encodes a pseudorabies virus gD protein. Further, the encoding element encodes gD wild-type protein, gD truncated protein, gD mutant protein and gD truncated mutant protein; preferred coding elements encode gD truncated plus muteins. Further, the encoding element encodes an optimized nucleotide sequence of pseudorabies virus gD truncated and mutant protein. Further, the circular RNA comprises an optimized translation initiation element. Researches show that the circular RNA provided by the invention can induce protein expression in an animal body, generate a high-level neutralizing antibody and successfully activate T cell immunity, especially CD8 + T cell immune response, and a basis is provided for preparing a circular mRNA vaccine for preventing and / or treating pseudorabies.

The invention discloses an mRNA (messenger ribonucleic acid) vaccine for preventing and / or treating pseudorabies and application of the mRNA vaccine. Codon optimization is performed on a gene sequence of a pseudorabies virus gD protein antigen to obtain an optimized gene sequence of the pseudorabies virus gD protein antigen; the pseudorabies virus truncated gD protein antigen is obtained by optimizing the sequence of the pseudorabies virus gD protein antigen, the gene sequence of the truncated gD protein antigen is subjected to codon optimization, and experiments show that the optimized pseudorabies virus truncated gD protein antigen has higher expression quantity; the sequence-optimized pseudorabies virus truncated gD protein is further prepared into an mRNA vaccine, and after the mRNA vaccine is used for immunizing mice and piglets, it is found that the mRNA vaccine of the sgD protein can play an immune protection role on the mice and the piglets, and the mRNA vaccine is obviously superior to commercially available pseudorabies inactivated vaccines.

The invention belongs to the technical field of antiviral drugs, and particularly relates to application of 18 beta glycyrrhetinic acid in preparation of drugs for resisting porcine pseudorabies virus infection. The invention provides application of 18 beta glycyrrhetinic acid in preparation of a medicine for resisting porcine pseudorabies virus infection in order to solve the problem that an existing PRV (porcine reproductive virus) new prevalent strain II type lacks vaccines and medicines capable of effectively protecting the existing PRV new prevalent strain II type. Meanwhile, the invention also provides a medicament containing 18 beta glycyrrhetinic acid for resisting porcine pseudorabies virus infection. It is found for the first time that the 18 beta glycyrrhetinic acid can inhibit adsorption and replication of PRV on PK-15 cells, and the 18 beta glycyrrhetinic acid has a very remarkable effect on resisting PRV, so that the 18 beta glycyrrhetinic acid can be used for preparing the medicine for resisting porcine pseudorabies virus infection, is safe, non-toxic and harmless, and provides an effective and reliable natural medicine for prevention, control and adjuvant treatment of porcine pseudorabies virus infection.

The invention belongs to the field of virus detection, and relates to a method for rapidly detecting porcine pseudorabies virus by RAA-LFD. The biological composition or the kit is provided, a sample to be detected reacts with the RAA primer group and the probe in an RAA-LFD reaction system, and the detection test strip is adopted to detect the reacted liquid. The RAA primer group is selected from any one of the following primer groups: a primer group of which the nucleotide sequences are respectively shown as SEQ ID NO.3 and SEQ ID NO.4, a primer group of which the nucleotide sequences are respectively shown as SEQ ID NO.5 and SEQ ID NO.6, or a primer group of which the nucleotide sequences are respectively shown as SEQ ID NO.15 and SEQ ID NO.16. Through combination of RAA and LFD, the kit can realize portability, sensitivity and high specificity, is suitable for on-site rapid detection of PRV, and provides technical support for prevention and control of PRV.

The invention discloses a dual fluorescent quantitative PCR (polymerase chain reaction) primer probe combination for identifying a PRV (pseudorabies virus) vaccine strain and a wild strain. The dual fluorescent quantitative PCR primer probe combination comprises two pairs of primers gB-F and gB-R, gE-F and gE-R, and probes gB-probe and gE-probe, wherein the two pairs of primers are used for specifically amplifying a PRV gB gene and a PRV gE gene; through existence of real-time fluorescence PCR amplification curves of the PRV gB gene and the gE gene, a PRV vaccine strain (gE gene deletion) and a wild strain can be rapidly, specifically, sensitively and stably identified, technical support can be provided for porcine pseudorabies epidemic disease monitoring, and thorough purification of the PRV in a live pig breeding link is facilitated.

Comprising a multivalent fusion protein and an exosome delivery carrier, and the content of the multivalent fusion protein is 70-90 [mu] g; wherein the multivalent fusion protein is a fusion protein formed by connecting a gB glycoprotein, a gC glycoprotein, a gD glycoprotein and a high-immunogenicity part of an important antigen segment selected from gE and gI; the exosome delivery carrier is a small vesicle extracted and purified from mesenchymal stem cells or HEK 293-F cells, the surface of the exosome delivery carrier is modified with a CD40L single-chain antibody, and the exosome delivery carrier is used for loading multivalent fusion protein; the porcine pseudorabies virus vaccine further comprises adjuvants, and the adjuvants comprise a TLR agonist, CpG oligonucleotide, synthetic liposome and a traditional aluminum salt adjuvant. The porcine pseudorabies virus vaccine also comprises a mucous membrane adsorbent, a penetration enhancer, a stabilizer and a preservative. The invention aims to construct a multivalent fusion protein complex containing a plurality of important antigen segments by using an advanced bioengineering technology, and provides a wider protection effect by combining an efficient exosome delivery platform and a method for enhancing immune response.

The invention relates to a porcine bacterial polysaccharide-viral protein conjugate vaccine as well as a preparation method and application thereof, and belongs to the technical field of animal vaccines. The vaccine takes porcine bacterial polysaccharide and virus protein as raw materials, and a polysaccharide-virus protein conjugate is formed through coupling of a biotin-avidin system; wherein the virus protein is selected from one of classical swine fever virus E2 protein, porcine circovirus type 2 Cap protein, porcine pseudorabies virus gD protein and the like, and the porcine bacterial polysaccharide is selected from one of streptococcus suis capsular polysaccharide, porcine pasteurella multocida capsular polysaccharide and the like. The vaccine can simultaneously induce an organism to generate a high-level IgG antibody aiming at porcine bacterium capsular polysaccharide and virus protein, so that synergistic immune protection on porcine bacterium and virus infection is realized, and a broad-spectrum cross immune effect is achieved.

The invention relates to the technical field of antibodies, and discloses an antibody for specifically recognizing porcine pseudorabies virus gB (507-734 aa) or an antigen binding fragment thereof and application. The antibody or the antigen binding fragment thereof comprises a heavy chain and a light chain; the antibody or the antigen binding fragment thereof can be used for detecting the existence or the level of the porcine pseudorabies virus gB protein in a sample, and has key significance on optimization of a porcine pseudorabies virus detection method, improvement of diagnosis accuracy and realization of effective prevention and control of epidemic situations.

The invention belongs to the technical field of biology, and particularly relates to a porcine pseudorabies virus gB-gD recombinant fusion protein subunit vaccine as well as a preparation method and application thereof. According to the invention, the PRV gB and gD genes are selected to serially express the gBgD protein to prepare the vaccine, the gB and gD proteins which are respectively expressed are replaced, and the vaccine is high in safety and good in immunogenicity, can effectively prevent the occurrence of a pseudorabies virus virulence enhancement event when being clinically used, provides a powerful space for purification of pseudorabies, and has a wide application prospect.

The present invention belongs to a new use of verrucine, and particularly relates to the use of verrucine and verrucine extract in the preparation of antiviral drugs. Verrucine has an inhibitory effect on porcine delta coronavirus, porcine epidemic diarrhea virus, pseudorabies virus, herpes simplex virus, influenza virus or African swine fever virus. The verrucine extract and verrucine of the present invention are very effective in antiviral treatment. The verrucine extract and verrucine of the present invention are very effective in broad-spectrum antiviral treatment, are safe, have few toxic and side effects, have low drug residues and are pollution-free.

This invention relates to an anti-swine pseudorabies virus (PRV) preparation and the application of Rehmannia glutinosa polysaccharide in its preparation, belonging to the field of biopharmaceutical technology. This invention provides an anti-swine pseudorabies virus (PRV) preparation comprising Rehmannia glutinosa polysaccharide. It also provides the application of Rehmannia glutinosa polysaccharide in the preparation of an anti-PRV preparation for the prevention and / or treatment of swine pseudorabies. This invention utilizes Rehmannia glutinosa polysaccharide to prevent infection by the PRV variant XJ5 by reducing intracellular oxidative stress levels; it also resists viral infection by affecting viral adsorption. Furthermore, when the concentration of Rehmannia glutinosa polysaccharide is between 50 and 400 μg / mL, it can also affect viral entry and replication, and has no cytotoxicity to PK-15 cells.

The invention relates to the technical field of biological medicines, and particularly discloses a diluent for a porcine pseudorabies live vaccine. The diluent for the porcine pseudorabies live vaccine is prepared by stirring and dissolving sodium dihydrogen phosphate, dipotassium phosphate, calcium chloride, sodium chloride, a traditional Chinese medicine extract, a stabilizer and water, and performing high-pressure steam sterilization, the traditional Chinese medicine extract is prepared by taking poria cocos and honeysuckle as traditional Chinese medicine raw materials and further reacting with chitosan and tannic acid, and the stabilizer is prepared by mixing gelatin, cyclodextrin and bovine serum albumin according to a mass ratio. When the diluent prepared by the method is used for diluting the porcine pseudorabies live vaccine, the stability and the activity of vaccine antigens can be effectively improved, the activity and the function of immune cells are enhanced, so that the immune effect of the vaccine is enhanced, and the prepared diluent for the porcine pseudorabies live vaccine solves the problem that a traditional live vaccine diluent does not have an immune enhancement effect.

The invention discloses application of PD-L1 as a drug target for resisting pseudorabies virus infection, the invention discloses PD-L1 as a key host limiting factor for pseudorabies virus infection for the first time, and knockout of PD-L1 can significantly promote replication of pseudorabies virus and induce apoptosis and inflammatory response of host cells. And PD-L1 overexpression can effectively inhibit proliferation of the pseudorabies virus and induce apoptosis and inflammatory response of host cells. Mechanism research results show that PD-L1 can be combined with envelope proteins gB and gD of the pseudorabies virus, and the effect of resisting pseudorabies virus infection is achieved through a cGAS-STNG-IFN pathway. The further research finds that the 125th arginine (ARG125) and the 271th lysine (LYS271) of the PD-L1 protein play a key role in the anti-pseudorabies virus infection effect of the PD-L1. The invention provides a new target spot for drugs for preventing or treating pseudorabies virus infection, and has a good application prospect in the aspect of resisting pseudorabies virus.

The invention relates to the technical field of biological medicines, and particularly discloses application of a swine STING molecular agonist SR-717 in preparation of a pseudorabies virus infection resisting medicine. The swine STING molecular agonist SR-717 can be stably combined with swine STING protein, the promoter activity of a downstream signal molecule interferon regulatory factor 3 (IRF3) and I-type interferon (IFN beta) is enhanced, PRV replication can be remarkably inhibited, the virus load is reduced, and pathological changes of infected mouse lung, brain and other tissues are reduced. The SR-717 provided by the invention has good targeting property and immune activation ability, and is suitable for intervention treatment of porcine pseudorabies virus infection.

The utility model relates to the technical field of farm disinfection, and disclose a pig pseudorabies prevention disinfection structure, including passageway frame, the upper surface fixed connection of passageway frame has the disinfection and killing main part mechanism, the output end fixed communication of disinfection and killing main part mechanism has the spray head, the outside of passageway frame is provided with non -contact disinfection mechanism, and non -contact disinfection mechanism includes two fixed frame, and the inside of every fixed frame all is provided with the shelter door pole. The pig pseudorabies prevention disinfection structure, through passageway frame, disinfection and killing main part mechanism, spray head, fixed frame, shelter door, button controller, warning light, warning device, first infrared distance sensor, second infrared distance sensor, servo motor and rotary lever, passageway frame can be installed at the pig house entrance and exit, and then when the breeding worker enters and exits passageway frame, first infrared distance sensor can monitor that personnel has stood in passageway frame, and second infrared distance sensor can monitor that there is no shelter at two shelter doors.

The present invention discloses a dual nano-PCR detection kit for porcine reproductive and respiratory syndrome virus and pseudorabies virus and its application, belonging to the technical field of molecular biology. The dual nano-PCR detection kit for porcine reproductive and respiratory syndrome virus and pseudorabies virus includes a primer set for detecting porcine reproductive and respiratory syndrome virus (PRRSV) and a primer set for detecting pseudorabies virus (PRV). The primers of the kit of the present invention have strong specificity, good repeatability and high sensitivity. Using nano-dual PCR for detection, it can detect as low as 5.17×10<supgt;2< / supgt> copies / μL and 4.46×10<supgt;2< / supgt> copies / μL, which is 100 times more sensitive than ordinary PCR; a single detection operation can simultaneously identify single infection or co-infection of the two viruses, greatly saving the detection time and reducing the detection cost, and providing technical support for disease prevention and control in pig farms.

This study describes the construction, identification, and application of a pseudorabies virus (PRV) triple-gene deletion vaccine strain (gI / gE / TK). Based on the JS-2012-ΔgI / gE double-gene deletion strain, the TK virulence gene was further deleted using CRISPR technology. Its pathogenicity was evaluated in KM mice and piglets, and its immunoprotective efficacy was verified in piglets. This provides theoretical support for the clinical prevention and control of pseudorabies; establishes experimental models in mice and pigs to study the immunoprotective efficacy of the triple-gene deletion strain; and prepares for the subsequent development and evaluation of vaccines using PRV strains as vectors.

This application relates to a neutralizing monoclonal antibody, hybridoma cell, and application thereof against pseudorabies virus gD protein. This application constructs a PRV gD eukaryotic expression vector, transduces it into CHO cells for expression, and then immunizes animals with PRV gD recombinant protein and a live attenuated vaccine as immunogens to produce a neutralizing monoclonal antibody against PRV gD. The neutralizing monoclonal antibody or hybridoma cell provided herein lays the foundation for the development of rapid detection technologies for pseudorabies virus and its gD protein. These antibodies have broad research and application value and commercial utility in pseudorabies virus-related immunoassays, and provide important guidance for the development of pseudorabies virus-based vaccines.

The invention belongs to the technical field of veterinary biological products, and particularly relates to a swine fever-porcine pseudorabies bivalent subunit vaccine containing a traditional Chinese medicine immunopotentiator as well as a preparation method and application of the swine fever-porcine pseudorabies bivalent subunit vaccine. The enhancer is prepared by compounding a hypericum perforatum extract and a horse chestnut seed extract, the hypericum perforatum extract and the horse chestnut seed extract are mixed with a classical swine fever virus (CSFV) E2 protein, a pseudorabies virus (PRV) gB protein, a PRV gD protein and an adjuvant to prepare a subunit vaccine, the vaccine can remarkably improve the humoral immunity level and the cellular immunity level excited by the vaccine at the same time, and the vaccine can be used for preparing the vaccine. Specifically, cell factors such as neutralizing antibodies GMT and IFN-gamma are remarkably increased, and the uniformity and stability of immune response are promoted, so that the immunogenicity of the existing swine fever-porcine pseudorabies bigeminy subunit vaccine is effectively improved, and the protection effect of the vaccine is enhanced.

The application discloses a recombinant pseudorabies virus strain expressing classical swine fever virus E2 protein and application, and relates to the technical field of biology.The recombinant E2 protein provided by the application successfully realizes high-level expression of the E2 protein by removing the transmembrane region of the E2 protein and using the 18aa signal peptide of the E2 protein itself and a pig albumin signal peptide.The recombinant PRV expressing the classical swine fever virus (CSFV) E2 protein can induce E2 antibodies and cellular immunity after immunization of mice.The recombinant pseudorabies virus strain provided by the application can be used as a bivalent vaccine for preventing CSFV and PRV infection.

The invention relates to the technical field of biological medicines, and aims to provide application of glutamic acid in preparation of a medicine for treating pseudorabies virus infection. Particularly, glutamic acid is applied to preparation of drugs for treating pseudorabies virus infection. The medicine takes glutamic acid as an active component, and comprises an effective amount of glutamic acid and pharmaceutically acceptable auxiliary materials. It is proved for the first time that glutamic acid can remarkably inhibit PRV replication in PK-15 cells, the effectiveness of glutamic acid in the aspects of inhibiting virus replication and improving infection symptoms and the remarkable effect of glutamic acid in the aspects of improving the survival rate and weight change of infected mice are verified, and a new scientific basis and practical support are provided for prevention and control of PRV infection. The traditional Chinese medicine composition has the characteristics of high safety and small toxic and side effects, and is low in price, easy to obtain and suitable for being popularized and used in farms. A novel treatment method is provided for coping with prevalence of porcine pseudorabies, virus transmission in pig farms is expected to be prevented, and economic losses are reduced.

This invention relates to the field of biomedical technology, specifically disclosing a STING agonist ST-71, its synthesis method, and its applications. ST-71 is obtained by modifying SR-717. The STING agonist ST-71 synthesized in this invention has the characteristics of good antiviral effect, low toxicity and side effects, and cost-effectiveness, and can also inhibit the proliferation of porcine pseudorabies virus.

The application discloses a reagent containing pig pseudorabies virus gB protein magnetic microparticle coating, a preparation method and a kit, and relates to the technical field of antibody detection. The preparation method of the reagent containing pig pseudorabies virus gB protein magnetic microparticle coating comprises the following steps: incubating magnetic microparticles and gB protein antibodies in a coating buffer solution; removing the reaction solution, and performing first sealing on the gB protein magnetic microparticle coating with a first sealing solution; after the first sealing is completed, the first sealing solution is washed away, and then the gB protein magnetic microparticle coating is sealed for the second time with a second sealing solution; after the second sealing is completed, the second sealing solution is washed away, and then resuspension is performed to obtain the reagent containing the gB protein magnetic microparticle coating. The two-step sealing method is adopted to prepare the reagent containing the pig pseudorabies virus gB protein magnetic microparticle coating, the sealing effect is good, and non-specific adsorption in the system is reduced. The kit provided by the application contains the reagent, and has the advantages of high analysis sensitivity, a wide linear range and high accuracy.

This invention relates to the field of biomedical technology, specifically to a stably expressed porcine pseudorabies virus gD recombinant protein, a vaccine composition, its preparation method, and its applications. The invention provides a stably expressed porcine pseudorabies virus gD recombinant protein, the amino acid sequence of which is shown in SEQ ID NO:7. The porcine pseudorabies virus gD recombinant protein is stably expressed in monoclonal cells, exhibiting stable expression levels after 48 consecutive passages. This lays a solid foundation for its subsequent applications in the preparation of stable porcine pseudorabies subunit vaccine compositions or in porcine pseudorabies virus vaccine efficacy testing formulations.

Provided is a method for constructing a gE gene-deleted pseudorabies virus (PRV) strain using an adenine base editor (ABE) and use thereof. The method includes: designing an sgRNA sequence using the ABE with a start codon of the gE gene in a PRV as a target site, ligating an enzyme-digested fragment to a double-stranded DNA fragment with sticky ends to obtain a ligation product; and transforming the ligation product into a competent cell to allow plate screening and culture, selecting a resulting positive bacterial strain to allow expanded culture, and extracting a plasmid from a resulting positive bacterial solution; and transferring the plasmid into a target cell to allow the transfection for 24 h, collecting a resulting virus liquid, and centrifuging the virus liquid to collect a resulting supernatant.