73 results about "Pseudorabies Vaccines" patented technology

The invention provides a Chinese herbal medicinal feed additive for preventing and controlling swine infectious disease. The swine feed additive consists of 37 Chinese medicinal herbs, namely, gypsum, rehmannia root, rhinoceros horn, golden thread, cape jasmine fruit, tree peony bark, baical skullcap root, red paeony root, figwort root, common anemarrhena rhizome, forsythia suspensa , platycodon root, liquorice root, common lophatherum herb, amur corktree bark, honeysuckle flower, Chinese pulsatilla root, indigowoad root, heartleaf houttuynia herb, astragalus, szechwon tangshan root, hawkthorn fruit, medicated leaven, barley sprout, radish seed, chicken's gizzard -membrane, Chinese thorowax root, common andrographis herb, philippine violet herb, tuber fleeceflower root, massa medicata fermentata fujianensis, cyrtomium rhizome, tung leaf, tangerine peel, white paeony root, pine needle and indigowoad leaf through scientific compatibility. The feed additive is added into swine feed in the proportion; under the condition of not using any vaccine, the feed additive can effectively prevent and cure severe mixed flu symptoms, infection and other syndromes caused by swine respiratory disease, asthma, contagious pleuropneumonia, swine virus mixed flu, high swine fever, porcine circovirus, swine fever, flu, pseudorabies, salmonellosis, bacillosis, streptococcus, erysipelas, paratyphoid, eperythrozoon, toxoplasm and other multi-pathogeny and provides genuine green food for the market.

The invention provides a heatproof lyoprotectant for a live vaccine against pseudorabies. The heatproof lyoprotectant comprises, by weight, 3 to 10 parts of gelatin, 1 to 5 parts of trehalose, 5 to 15 parts of sucrose, 0.1 to 2 parts bovine serum albumin, 1 to 8 parts of tryptone, 2 to 10 parts of enzyme-hydrolyzed casein, 1 to 5 parts of thiourea, 0.8 to 2 parts of L-monosodium glutamate, 0.1 to 3 parts of arginine, 0.5 to 5 parts of polyvinylpyrrolidone (PVP-K30) and 0.1 to 2 parts of mannitol. The invention also discloses a preparation method of the heatproof lyoprotectant and a lyophilized vaccine prepared from the heatproof lyoprotectant. When the heatproof lyoprotectant is used for protecting the vaccine and a specific lyophilization process is employed, lyophilization loss of viruses can be effectively reduced, the temperature tolerance of the viruses can be improved, and the vaccine can still maintain good physical properties and titer after long-term storage; i.e., the vaccine has stable characters and has the characteristics of heat resistance and long storage time.

The invention discloses a pseudorabies virus gene-deleted attenuated strain. The deleted gene sequences of the attenuated strain include a nucleotide sequence for coding No.198 to No.366 amino acids of gI protein of a pseudorabies virus and a nucleotide sequence for coding No.1 to No.404 amino acids of gE protein of the pseudorabies virus. The invention further discloses a preparation method and an application of the pseudorabies virus gene-deleted attenuated strain. The pseudorabies virus gene-deleted attenuated strain has a good immune protection effect on the pseudorabies virus and can be used as a vaccine candidate strain for preventing and treating pseudorabies.

The invention discloses a porcine pseudorabies virus positive serum preparation method. A water-in-oil type inactivated vaccine is prepared through emulsification, and then porcine pseudorabies virus positive serum with a high neutralizing titer is obtained through immune reaction generated by stimulating sheep through the synergistic effect of a porcine pseudorabies virus live vaccine and the inactivated vaccine. The method reduces the risk of exogenous virus contamination caused by homologous animals when positive serum is prepared from pigs, and a new thought is provide for preparation of positive serum.

The invention relates to the technical field of swine pseudorabies viruses, and relates to a swine pseudorabies virus variant XF-1 strain, a preparing method thereof and applications of the strain. The accession number of the variant is CCTCC NO V201654. The swine pseudorabies virus variant (the XF-1 strain) is adopted to prepare an inactivated vaccine, and safety and immunoprotective experiments are performed. Results prove that the swine pseudorabies virus variant (the XF-1 strain) inactivated vaccine has good safety, and that the immunoprotection efficiency of the inactivated vaccine is obviously higher than that of other immune groups, and therefore the virus strain is proved to have good immunogenicity, can be used for vaccine research and development for the swine pseudorabies, and has a food vaccine development prospect.

The invention relates to a variable region sequence of a mouse monoclonal antibody which specifically binds to a pseudorabies virus gD protein. The sequence is the variable region sequence of mouse monoclonal antibody 3B6 or the variable region sequence of mouse monoclonal antibody 5G7. The invention also relates to an antibody consisting of a heavy chain variable region sequence or its conservative variant and / or a light chain variable region sequence or its conservative variant of mouse monoclonal antibody 3B6, and an antibody consisting of a heavy chain variable region sequence or its conservative variant and / or a light chain variable region sequence or its conservative variant of mouse monoclonal antibody 5G7. According to the invention, an ELISA detection kit containing the antibody overcomes the technical problem that the PRVgD protein cannot be accurately quantified after tandem expression or fusion expression with other proteins and the expected expression effect cannot be verified, and the key problem of formulating a vaccine is solved. Meanwhile, a pharmaceutical composition containing the antibody has a wide spectrum.

The invention relates to a formula of a medicament for treating swine high fever and a using method thereof. The medicament is prepared by dissolving 0.5 to 25 percent of Chinese angelica extract, 0.5 to 25 percent of rhubarb extract, 0.5 to 25 percent of Himalayan teasel root extract, 0.5 to 25 percent of common clubmoss herb extract, 0.5 to 25 percent of liquorice root extract, 0.5 to 25 percent of croton seed extract, 0.5 to 25 percent of toad venom, 0.5 to 25 percent of cicada slough extract, 0.5 to 25 percent of radix scutellariae extract, 0.5 to 25 percent of honeysuckle extract and 0.5 to 25 percent of common andrographis herb extract with solvent normal saline to obtain an injection, wherein the sum of the components and the balance is one hundred percent. The medicament has special effects on early and middle high fever, fulminant epidemic diseases from 2006 to 2009, mixed epidemic swine fever blue-eared disease of virus, pathogenic bacteria and worm, pseudorabies, blood worm, toxoplasmosis, lung plague, erysipelas, eircovims, streptococcus, haemophilus diseases, pneumonia, bronchitis and abdominal respiration and can relieve nose and mouth foaming in two hours.

The invention discloses a traditional Chinese medicine combination for treating porcine pseudorabies and a preparation method and application thereof. The combination is prepared with 15 to 30 parts by weight of sheepear inula herb, 6 to 18 parts by weight of trametes robiniophila murr and 5 to 15 parts by weight of fiveleaf gynostemma herb. The preparation method for the combination includes thefollowing steps: the sheepear inula herb, the trametes robiniophila murr and the fiveleaf gynostemma herb are decocted for 2 to 3 times; the decoctions are combined, concentrated to be thick and mixed with ethanol until the ethanol concentration is 60 to 70 percent; after deposition and filtration, the concentrated liquor is then mixed with ethanol until the ethanol concentration is 75 to 80 percent; after deposition, filtration and filtrate concentration, the liquor is added with distilled water and refrigerated for deposition; and after filtration and concentration, the traditional Chinese medicine combination is obtained. When being used for treating porcine pseudorabies, the traditional Chinese medicine combination has little side effects and a high curative rate, and is safe and convenient to use.

The invention discloses the process for preparing anti-Pseudorabies yolk (igY) which comprises, (1) employing pig's hydrophobia vaccine or pig's hydrophobia genetic engineering deletion vaccine purifying grade testing egg chickens, carrying out potency determination through enzyme linked immunoadsorption test, collecting immunological eggs, (2) reinforcing the immunological vitelline, carrying acid treatment through water dilution and lactic acid regulation, so as to remove impure proteins, carrying out phthalate methyl hydroxypropylcellulose treatment and freeze melting treatment to remove grease, desalinizing the filter liquor through ammonium sulfate salting out and dialysis, finally carrying out column chromatography to obtain the high purity anti-Pseudorabies igY with high sensitivity and strong specificity.

The invention discloses a breeding method for breeding pigs, and relates to the technical field of pig breeding. In the breeding pig breeding process, the breeding pigs in different phases such as piglet phase, reserve swine phase and sow phase are subjected to immunization by various vaccines in different time stages. According to the breeding method, a normative immunization program is provided, so that swine fever, foot-and-mouth diseases, pseudorabies, porcine reproductive and respiratory syndrome, parvovirus, epidemic diarrhea, contagious gastroenteritis porcine rotavirus, epidemic encephalitis B, haemophilus parasuis, porcine circovirus and the like which are common in the piglets, the reserve swines and the sows can be effectively prevented.

The invention provides a compound propolis composition for treating porcine pseudorabies and a preparation method thereof, and relates to the field of animal medicines. The preparation method provided by the invention comprises the following steps: (1) micronizing each component except interferon in the prescription respectively, and screening through a 700-mesh screen; (2) weighing 15 to 35 parts of propolis, 10 to 20 parts of porcine interferon, 15 to 45 parts of astragalus mongholicus, 15 to 35 parts of isatis root and 10 to 45 parts of gastrodia elata according to weight proportion, and mixing for 5 to 10 minutes until the mixture is uniform; (3) weighing 10 to 35 parts of humifuse euphorbia herb, 10 to 35 parts of houttuynia cordata, 15 to 35 parts of barbed skull cap herb, 15 to 35 parts of fructus forsythiae and 15 to 35 parts of toad according to the weight proportion, and mixing for 5 to 10 minutes until the mixture is uniform; and (4) combining compositions prepared in step (2) and step (3), and continuing mixing for 10 to 15 minutes until the mixture is uniform. The compound propolis composition provided by the invention has the advantages that: the components are reasonable, and the preparation is easy; the composition has no toxic or side effect, no pollution, no medicinal residues, no drug resistance, micronization, high bioavailability and remarkable curative effect; and the clinical application proves that the composition has special effect in treating porcine pseudorabies.

The invention provides a pseudorabies virus (PRV) digene deletion attenuated strain and application thereof, and belongs to the field of vaccines for animal medicine. A preservation number of a PRV LA1206-80 strain is CGMCC (China General Microbiological Culture Collection Center) No.14329. The invention also provides application of the PRV digene deletion attenuated strain in preparation of a pseudorabies vaccine, and a recombinant vector live vaccine. A recombinant vector is obtained by inserting an exogenous protective antigen gene expression cassette into an attenuated strain genome. The live vaccine of the PRV LA1206-80 strain is absolutely safe to one-day-old neonatal piglets, can be used for early immunization of the neonatal piglets and blocks off infection of wild type viruses asearly as possible. The vaccine is inoculated to weaned PRV negative piglets, so that a high-efficiency protecting force can be quickly generated; particularly, the vaccine can prevent attacking and detoxify. By monitoring PRV gE and gB antibodies, purification on a PRV variant strain is greatly facilitated.

The invention discloses a triple fluorescent quantitative PCR primer, probe and kit for identifying pseudorabies virus classical strains, variant strains and Bartha-K61 vaccine strains. The sequences of the primer are represented by SEQ ID No. 8-9 and SEQ ID No. 14-15, and the sequences of the probe are represented by SEQ ID No. 1, 4 and 6. According to the kit, the pseudorabies virus classical strains, the variant strains and the Bartha-K61 vaccine strains can be accurately identified based on TaqMan probes marked with different fluorescence signals, and the kit has good sensitivity and specificity and can be applied to the discriminative detection of pseudorabies virus strains.

The invention discloses primers and a probe set of a cross primer amplification-immunochromatography test paper combination method for detecting a pseudorabies virus wild strain and a cross primer amplification-immunochromatography test paper combination kit for detecting the pseudorabies virus wild strain. The invention first discloses the primers and the probe set of the cross primer amplification-immunochromatography test paper combination method for detecting the pseudorabies virus wild strain, which comprise two outer side primers, a cross primer and two probes. The invention further discloses the cross primer amplification-immunochromatography test paper combination kit for detecting the pseudorabies virus wild strain, which comprises a cross primer amplification agent and an immunochromatographic test strip. Virus genomic DNA is taken as a template, a CPA reaction system is built through the cross primer amplification agent and is used for amplification, and a sample of an amplified product is loaded onto the immunochromatographic test strip, so as to realize result determination. The primers, the probe set and the kit, provided by the invention, can realize specific detection on the pseudorabies virus wild strain, the sensitivity and specificity are good, the visual degree is high, the operation is simple and convenient, and the primers, the probe set and the kit are suitable for on-site detection of pseudorabies viruses.

The invention relates to the field of viruses, in particular to a porcine pseudorabies double gene deletion mutant virus strain and an establishment method thereof. The virus strain is a gE, gI double gene deletion strain, the preservation number is CCTCC NO. V201749, preservation unit is: China's typical culture collection center, the preservation address is: Wuhan University, Wuhan, China, the postcode is 430072. rPRV HN2012-gE- / gI-immunized piglets established do not have any adverse reactions during the experiment, the neutralizing antibody level verifies that the deletion virus maintains good immunogenicity, the obtained rPRV HN2012-gE- / gI-double gene deletion virus is safe for the piglets, PRV (pseudorabies virus) specific antibody can be produced effectively and rapidly after immunization, and the porcine pseudorabies double gene deletion mutant virus strain is a vaccine strain which is expected to be used in the prevention and control of PRV epidemic.

The invention provides a novel nucleic acid amplification detection method for synchronously and rapidly detecting three important pig pathogeny of porcine circovirus type 2, porcine pseudorabies virus and porcine parvovirus by LNA (Locked nucleic acid) modified base common primer guidance as well as a detection kit. A pair of common primers containing LNA modified bases and three pairs of 5'end virus specific primers with common primer base sequences are designed and screened, and the specific PCR (Polymerase Chain Reaction) amplification reaction is guided mainly by the LNA modified base common primers in the reaction process by an optimized reaction system and two steps of PCR amplification reaction conditions, so that the porcine circovirus type 2, porcine pseudorabies virus and porcine parvovirus are detected simply, conveniently and synchronously. The novel nucleic acid amplification detection method and the kit which are provided by the invention verify that the LNA modified base common primers can efficiently guide multiple PCR amplification, and provide research and experiment basis for the research and the invention of the modified base common primers applicable to multiple PCR systems with other functions.

The invention discloses a bivalent gold-labeled test strip for a classical swine fevervirus and a porcine pseudorabies virus and a preparation method thereof. The bivalent gold-labeled test strip comprises a support plate, a sample pad, a gold label pad, an NC membrane and an absorbent pad; the NC membrane contains the imprints of a classical swine fevervirus detection line T1 ''|'', a porcine pseudorabies virus detection line T2 ''|'' and a quality control line C ''|''. During detection, when three red strips ''|||'' appear on a detection membrane, the test strip shows that there is the co-infection of the classical swine fevervirus and porcine pseudorabies virus; when two red strips ''||'' of the detection line T1 and the quality control line C appear, the test strip shows that there isonly the swine fever virus infection; when two red strips ''||'' of the detection line T2 and the quality control line C appear, the test strip shows that there is only the porcine pseudorabies virusinfection; and when only one red strip ''|'' appears, the test strip shows that the result is negative, and both viruses are not infected. The test strip is strong in specificity, high in sensitivity,wide in response spectrum, can detect most existing epidemic isolates, simple and rapid in operation, can be widely used for the detection of classical swine fevervirus infection and porcine pseudorabies virus infection, and is easy to be popularized and applied in production practice.

The invention discloses a detection kit and a detection method for a porcine pseudorabies gB antibody. The kit detects the content of the antibody in a sample to be detected through a specific reaction of a magnetic particle-coated antigen, a luminescent marker-labeled protein and the antibody in the sample to be detected. The kit realizes the quantitative detection of the porcine pseudorabies gBantibody, and has the advantages of high sensitivity, wide detection range, fastness, good repeatability, and realization of fully automatic operation and high-throughput detection.

The invention discloses a pseudorabies virus gene engineering gB recombinant attenuated vaccine strain and application thereof. CRISPR / Cas9 is combined with techniques such as homologous recombination, and a genome of a pseudorabies virus vaccine strain Bartha-K61 is taken as a framework, and a recombinant pseudorabies virus strain Bar-JS-gB (BJB) is successfully acquired by replacing a gB gene of the Bartha-K61 with a gB gene of an epidemic pseudorabies virus variant JS-2012 in China. The strain BJB has stable heredity, and an immune mouse processed by an inactivated vaccine prepared by taking the strain BJB as a vaccine strain has relatively good capacity of effectively resisting attacking of a virulent strain JS-2012 than an immune mouse processed by an inactivated vaccine prepared from the Bartha-K61. Therefore, the strain BJB has relatively good immunogenicity, can be used for effectively controlling the prevalence of a PRV variant in China and can play an important role on control of pseudorabies.

The present invention relates to variable region sequences of a murine monoclonal antibody that specifically binds to a pseudorabies virus gC protein. The variable region sequences are variable regionsequences of murine monoclonal antibody 1B9 or variable region sequences of murine monoclonal antibody 1F2. The invention also relates to a pseudorabies virus gC protein antibody consisting of heavychain variable region sequences in the variable region sequences, or conservative variants thereof, and / or corresponding light chain variable region sequences in the variable region sequences, or conservative variants thereof. According to the immunohistochemical method established by the antibody, the problems can be solved that PCR cannot be clearly positioned in tissues and cells, and the dynamic distribution, qualitative and semi-quantitative detection of pathogenic organisms in a body cannot be tracked. The kit containing the antibody is used for accurately detecting the content of the PRVgC protein based on the dual monoclonal antibody sandwich principle, thereby solving the technical problems that the PRVgC protein cannot be accurately quantified after being expressed or fused in series with other proteins, and the expression effect cannot be verified.

The invention discloses a low serum culture medium for BHK-21 cell culture and corresponding virus production. The low serum culture medium is prepared from the following components: amino acid, a vitamin, inorganic salts, a protein, microelements and auxiliary components; the amino acid comprises glycine, L-arginine monohydrochloride, L-asparaginate, L-glutamic acid, L-glutamine, L-lysine hydrochloride and L-serine; and the protein comprises 4-HEPES, sodium dihydrogen phosphate monohydrate, milk protein hydrolysate, bovine serum albumin and recombinant human insulin. According to the low serum culture medium for the BHK-21 cell culture and the corresponding virus production, serum is no longer needed to be added in the maintenance stage of a pseudorabies vaccine produced from a BHK-21 cell, dosage of the serum is further reduced, the good growth state of cells can still be maintained, virus titer is improved, reduction of poison value of a semi-finished product is small, and preservation time of the semi-finished product is prolonged.

The invention relates to a traditional Chinese medicine composition for treating porcine pseudorabies. The traditional Chinese medicine composition is prepared from the following mixed components in parts by weight: 15-35 parts of puffball, 10-20 parts of semen strychni, 15-45 parts of vervain, 15-35 parts of cowherb seed, 15-45 parts of henbane, 10-35 parts of licorice, 10-35 parts of herba aristolochiae, 15-35 parts of radices trichosanthis, 15-35 parts of tabasheer and 15-35 parts of processed rhizoma arisaematis. The traditional Chinese medicine composition has the beneficial effects that the components are reasonable, the preparation method is simple, the composition does not have toxic and side effects, pollution, drug residue and drug resistance, is in superfine grinding, high in bioavailability and very obvious in curative effect and is clinically proved to have a special effect for treating porcine pseudorabies.

The invention discloses a visible gene chip for a pig pseudorabies virus, porcine parvovirus and porcine circovirus type-II. The visible gene chip comprises a solid phase carrier and a probe fixed on the solid phase carrier, wherein the probe comprises gene segments as shown in SEQ ID NO:1, 3, and 4; and the gene segments are used for respectively detecting the pig pseudorabies virus, the porcine parvovirus and the porcine circovirus type-II. The invention further discloses a kit for detecting the pig pseudorabies virus, the porcine parvovirus and the porcine circovirus type-II. By adopting the gene chip and the kit disclosed by the invention, pathogens of the pig pseudorabies virus, the porcine parvovirus and the porcine circovirus type-II can be effectively detected, infection caused by vaccine strains and wild strains of the pig pseudorabies virus can be distinguished, the gene chip and the kit have the characteristics of good specificity, high sensitivity and long preservation period, are short in time, rapid in detection, visible in detection result observation and good in clinical application prospect, no expensive equipment is needed, and the infection situation of livestock diseases can be rapidly handled.

The present invention provides recombinant DNA molecules comprising a sequence encoding a pseudorabies virus (PRV) glycoprotein selected from the group consisting of gI, gp50, and gp63, host cells transformed by said recombinant DNA molecule sequences, the gI, gp50 and gp63 polypeptides. The present invention also provides subunit vaccines for PRV, methods for protecting animals against PRV infection and methods for distinguishing between infected and vaccinated animals.

The invention relates to a variable region sequence of a mouse monoclonal antibody which specifically binds to a pseudorabies virus gB protein. The sequence is the variable region sequence of mouse monoclonal antibody 3C12 or the variable region sequence of mouse monoclonal antibody 5G12. The invention also relates to an antibody consisting of a heavy chain variable region sequence or its conservative variant and / or a light chain variable region sequence or its conservative variant of mouse monoclonal antibody 3C12, and an antibody consisting of a heavy chain variable region sequence or its conservative variant and / or a light chain variable region sequence or its conservative variant of mouse monoclonal antibody 5G12. According to the invention, an ELISA detection kit containing the antibody overcomes the technical problem that the PRVgB protein cannot be accurately quantified after tandem expression or fusion expression with other proteins and the expected expression effect cannot beverified, and the key problem of formulating a vaccine is solved. Meanwhile, the antibody can be used for immunohistochemical detection.

The invention provides a preparation method of high-potency positive serum for a porcine pseudorabies virus. The method takes a rabbit as an immunization object, and immunizes the rabbit with an inactivated vaccine containing a suitable amount of PolyI:C, muramyl dipeptide and cimetidine to produce the hyperimmune serum, can achieve the effect that no cytopathic effect occurs in subculture of at least 2 generations in a T25 square flask equal volume ratio neutralization 1 vaccine experiment, and meets the requirement that a cell method serves as an exogenous virus detection standard in the <Chinese veterinary pharmacopoeia> (2015 edition). The preparation method of the high-potency positive serum for the porcine pseudorabies virus has a good application prospect.

The invention provides a pseudorabies virus GE protein and preparation of a colloidal-gold immunochromatographic strip thereof. The pseudorabies virus colloidal-gold immunochromatographic strip is provided with a carrier plate, a sample pad, a colloidal-gold conjugate pad, a laminate membrane, a detection line, a quality-control line and an absorption pad. The expressed recombinant protein existedin a supernatant is easy to purify. The detection method established by taking the recombinant protein, which is purified by affinity chromatography, as a diagnostic antigen is low in cost and simpleand convenient, and especially the diagnostic antigen concentrates major antigen-epitopes of the gE antigen. The detection sensitivity and specificity of the recombinant protein are both higher thanthe intact gE protein.

The invention relates to a preparation method of a swine pseudorabies virus high-immune serum. The serum is obtained by taking pigs as immunization objects, conducting primary basic immunization and secondary enhanced immunization by using an inactivated vaccine, primarily counteracting toxic substances by using a porcine pseudorabies virus, and then executing collection; the inactivated vaccine contains an immunopotentiator of interferon alpha, interferon beta, astragalus polysaccharide and a macrophage colony stimulating factor. The swine pseudorabies virus high-immune serum can overcome theshortcomings of poor neutralization capacity of high-immune serum antibodies derived from pigs, and can be widely applied to strains JS-2012, Bartha-k61, HB-98, HB2000 and / or JS-A1 of pseudorabies viruses and has a good application prospect.

The invention discloses a crossing isothermal amplification primer set for detecting a pseudorabies virus wild strain, a kit and application thereof. The primer set comprises the primer set with the nucleotide sequences shown as SEQ ID NO.1-5. The kit comprises the primer set and a nucleic acid test strip. The use method of the kit comprises the following steps: firstly, preparing a crossing primer amplified reaction system, detecting a product acquired through thermostatic reaction with the nucleic acid test strip and then directly reading: wherein two strips indicate a positive result, one strip is located in a detection area and the other strip is located in a quality control area. The kit has the advantages of simple operation, low cost, easily observed reaction result, excellent specificity, suitability for on-site detection for export quarantine, food hygiene and livestock farm and easiness in wide application and popularization.