163 results about "Duck viral hepatitis" patented technology

The invention relates to an enzyme-linked immunosorbent assay (ELISA) kit for duck hepatitis virus type-I serum antibody and relates to a test method and application of the kit. The kit comprises an enzyme label plate coated by the recombinant VP1 (virus protein) protein, a rabbit anti-duck IgY antibody marked by horseradish peroxidase, a TMB substrate colour reagent, a positive serum, a negative serum and a kit specification. In the invention, by adopting the polymerase chain reaction, the VP1 genes are amplified from the DHV-1genome and the VP1 gene-containing recombinant expression plasmid pET32a-VP1 is constructed; the plasmid is transferred to host cells BL21 (DE3), and the in-vitro expression VP1 protein is purified by a nickel column and then used as the antigen; the enzyme-linked immunosorbent assay kit is established; the positive serum is the standard positive serum of duck hepatitis virus type-I and the negative control is the standard negative serum of duck. The test kit has the advantages of strong specificity, high sensitivity, simple operation, easy large-scale popularization and application, very important application value in diagnosis of duck hepatitis virus type-I, survey of epidemiology and immunization survey and the like.

The invention relates to a method for preparing a duck virus hepatitis divalent refined egg yolk antibody. The method is that vaccines of a duck virus hepatitis virus 1 type YBH1 strain and novel YBHX strain with good immunogenicity are selected from duck virus hepatitis superior prevalent strains to prepare strains, the strains are respectively inoculated to a chicken embryo and a duck embryo, a dead embryo body and the embryo juice are respectively obtained, the virus liquid is collected after grinding and freeze thawing, ultrafiltration concentration and inactivation with formaldehyde solution are carried out, and oil adjuvant is added to mix and emulsify to produce the vaccine. The vaccine is inoculated to a laying hen, the egg with high immunity is obtained, the yolk is separated, inactivated, extracted and refined to produce the duck virus hepatitis divalent refined egg yolk antibody which can prevent the duck virus hepatitis which is caused by the 1 type and novel duck hepatitis viruses, so the egg yolk has the advantages of high efficiency, good safety, high protection ratio and the like.

The invention provides a method for preparing a duck virus hepatitis refined heterogenous yolk antibody, comprising the following specific steps: (1) separating the yolk of a high-immunity egg; (2) extracting a yolk antibody; and (3) preparing a preparation and packaging. The method for preparing the duck virus hepatitis refined heterogenous yolk antibody is easy to operate and is applicable to mass production, and the prepared duck virus hepatitis yolk antibody has the advantages of stable property, high purity, high valence, stronger specificity and no drug residue, is mild and environmentally friendly and has application value in preventing and treating duck virus hepatitis, thus a new way is provided for preparing the duck virus hepatitis refined heterogenous yolk antibody.

The invention discloses a method for preparing an anti-duck viral hepatitis transfer factor, which comprises the following steps of: firstly, inoculating duck viral hepatitis in a chick embryo allantoic cavity for multiplication and extracting a virus antigen from the chick embryo allantoic cavity; secondly, immunizing a healthy pig body with the extracted virus antigen; and thirdly, extracting the anti-duck viral hepatitis transfer factor from the spleen of the immune pig. In the invention, specific transfer factor is extracted from duck viral hepatitis viruses and can effectively suppress the duck viral hepatitis viruses and protect the cells of a normal body from being infected with the duck viral hepatitis viruses, so that the duck viral hepatitis is prevented and treated radically. Moreover, the preparation method of the invention is simple and feasible, short in production time and low in cost and has a promising application prospect.

The invention relates to a duck virus hepatitis yolk antibody freeze-dried powder which comprises the following components: 85-88% of duck virus hepatitis yolk antibody, 0.8-1.0% of octanoic acid, 0.01-0.02% of thimerosal, 0.2-0.3% of formaldehyde, 4-5% of glucose, 1-2% of sorbic alcohol, 2-3% of glycine and 2-3% of mannitol. The freeze-dried powder has the advantages of long preservation time, high dissolving speed, high antibody titer and excellent biological activity, and more importantly, the purity of the antibody of the product is higher than that of the existing product on the market. At the same time, the invention also discloses a preparation method of the duck virus hepatitis yolk antibody freeze-dried powder, comprising the following steps: voluntarily separating strains in a disease prevalence area to prepare a vaccine, immunizing healthy dusks to obtain hyper-immune eggs, ad performing disinfection, acidification, octanoic acid treatment, refining extraction and purification, preparation, freezing and the like on the hyper-immune eggs to obtain the duck virus hepatitis yolk antibody freeze-dried powder. According to the preparation method in the application, a yolk solution is heated at the temperature of pasteurization when the acidification water treatment is carried out, so that not only is the disinfection purpose fulfilled, but also the recovery rate of the antibody and the clarity of the solution in the acidification treatment are increased, the prepared yolk antibody has the advantages of high biological activity, long antibody preservation time, titer reduction, lowered speed and the like.

The invention discloses a lentogenic CH60 strain of duck virual hepatitis virus. The lentogenic CH60 strain of the duck virual hepatitis virus is preserved in the China Center for Type Culture Collection of Wuhan University in China; and the preservation number is CCTCC NO: V201248. As the strain of duck virus hepatitis attenuated vaccine, the lentogenic CH60 strain of duck virual hepatitis virus provided by the invention is good in security and good in immunogenicity.

The invention discloses methods for preparing a duck virus hepatitis strain and an inactivated vaccine. The strain is obtained with the method comprising the following steps of: collecting duck hepatitis material with obvious pathological changes in a duck farm where duck virus hepatitis prevails, washing with bi-anti-sterilization PBS, diluting and grinding, freeze thawing twice and performing centrifugal separation; diluting virus separated by sterilization PBS, vaccinating chick embryo, selecting chick embryo with classic pathological changes that is dead 48-96h after vaccination, respectively gaining chick embryo liquid, transferring for 18-25 generations continuously to obtain a duck virus hepatitis virus chick embryo suitable strain. The strain can be in production, vaccinated in the chick embryo to gain chick embryo liquid, and is emulsified after vaccination to prepare the inactivated vaccine. The inactivated vaccine is high in safety, has good immune efficacy to young ducks, long immune time for breeding ducks and high level of antibody of breeding ducks, and the young ducks bred by the breeding ducks can resist duck virus hepatitis strains.

The invention discloses a duck egg yolk antibody for preventing and treating virus hepatitis of ducklings, and a preparation method thereof. The duck egg yolk antibody is extracted from the primiparous eggs of a high-immunity homebred breed duck starting laying for one month. The preparation method comprises the following steps of: (a) highly immunizing the breed duck before laying; (b) collecting high-immunity primiparous breed duck eggs; (c) cleaning and sterilizing; (d) separating egg yolks; (e) diluting; (f) stirring and filtering; and (g) packaging and preserving. After the egg yolks are continuously diluted for 2 times by using normal saline, the DHV (Duck Hepatitis Virus) precipitating antibody is detected by a conventional method, the antibody titer is more than 1:64, and the HI titer of H5 is more than 1:512. 0.5mL of the duck egg yolk antibody is subcutaneously injected to a 3-day-old duckling for preventing virus hepatitis; and 1mL of egg yolk antibody to which antibiotics are added is subcutaneously injected to a duck for treating virus hepatitis, and the egg yolk antibody is re-injected to a seriously diseased duck every other day.

The invention discloses a method for preparing a bivalent yolk antibody for preventing and treating duck virus hepatitis I and III. The bivalent yolk antibody can be used for effectively preventing and treating the duck virus hepatitis I and III single and mixed infection and solving the problems that the existing vaccines, serum antibodies and yolk antibodies are not effective to the mixed infection. In addition, the bivalent yolk antibody is extracted by adopting a caprylic acid precipitation method. The method is safe, low in cost and suitable for mass production. The prepared bivalent yolk antibody has high purity and high titer, is stable in nature and is easy for storage.

The present invention is the preparation process of yolk antibody for preventing and treating viral hepatitis of green duck. The present invention prepares the yolk antibody for preventing and treating viral hepatitis of green duck with chicken as one heterogenic animal. That is, after inactivated oil emulsion vaccine of viral hepatitis of green duck is used to immunizing laying fowl, specific immunoglobulin is extracted from the yolk for preventing and treating viral hepatitis of green duck. Clinical test and use shows that the yolk antibody, when used in preventing and treating viral hepatitis of green duck, has high preventing and treating effect, long effective period and no risk of propagating diseases vertically of homological antibody.

The invention relates to the field of animal medicine, in particular to a method for detecting duck plague virus antibody. The method particularly comprises the steps of the preparation of solid phase antigen, primary antibody combination, secondary antibody combination, developing, detection, judgment and the like. The method is an indirect enzyme-linked immuno sorbent assay (ELISA) method established based on purified recombinant UL51 protein, which has good specificity, and when duck hepatitis virus (DHV), Riemerella anatipestifer (RA), duck Escherichia coli (E.coli) are detected, the detection results are all positive; intra batch or batch-to-batch repeated tests show that variation coefficients are all less than 10%, and duck plague virus (DPV) vaccine immunized duck positive sera diluted at ratio of 1:3200 can be detected.

The invention relates to duck virus hepatitis inactivated vaccine and a preparation method thereof. The preparation method comprises the following steps: taking a duck hepatitis virus virulent strain CGMCC No.3202 with excellent immunogenicity and obtained by the inventor through field separation as a viral strain for producing vaccine; inoculating the viral strain in a susceptible chick embryo and obtaining the embryo fluid; and carrying out emulsification after inactivation to prepare the safe effective duck virus hepatitis vaccine. The duck virus hepatitis inactivated vaccine aims to prevent duck virus hepatitis virus infection and fills a gap in the market, thereby providing conditions for controlling the spread of duck virus hepatitis on poultry farms.

The invention discloses a novel DHAV (duck hepatitis A virus) JS strain and application of the DHAV JS strain in duck virus hepatitis prevention and cure. The DHAV JS strain is obtained through the steps of separation culture, duck embryo neutralization test, PCR (polymerase chain reaction) detection of viruses and sequencing detection experiment of the viruses, and the strain is determined to be the DHAV serotype 3; and the microbial preservation number is CGMCCNo.6852. According to the invention, through taking the novel DHAV JS strain as the virus strain for producing vaccine, the novel DHAV inactivated vaccine is prepared through the steps of inoculating 10-day-old SPF (specific pathogen free) duck embryos, selecting the embryos died after incubating for 48 hours, collecting the embryo liquid, inactivating and emulsifying. The clinical application proves that the prepared inactivated vaccine is favorable in safety and conducive to prevention and control of the novel DHAV disease, can prevent the infection and outbreak of the novel DHAV in a targeted way, and can be in mass production and clinical application.

The present invention discloses a combination of Chinese medicines for poultry and the preparation method and application. Hypericum forsythia and bidens grass are distilled. And then distilled liquid and residua are collected. The residua and the residual liquid after distillation being added with a hepatitis grass and a climbing nightshade are decocted three times. The decoction liquid is combined and concentrated to a thick concentration. A 95 percent ethanol is diluted to alcohol concentration of 60 percent to 70 percent. The drug liquid after sedimentation, filtration and concentration added with 95 percent ethanol are transferred to alcohol concentration of 75 percent to 80 percent and then are sedimentated and filtrated. The filtrate is concentrated. The drug liquid added with distilled water is frozenly precipitated, filtrated and concentrated. The combination of Chinese medicines is produced out after a combination of the distilled liquid of the hypericum forsythia and the bidens and the decoction liquid of the hepatitis grass and the climbing nightshade are combined. The combination of Chinese medicines has a cure rate of about 90% for a viruses and bacterial infectious disease of poultry, can be used to prepare the animal drug for treating duck viral hepatitis and chicken newcastle disease and has little side effect, safe use and fast absorption.

A veterinary Chinese medicine for treating the viral hepatitis of duck is prepared from oriental wormwood, capejasmine fruit and rhubarb in weight ratio of (1-6): (1-4): (1-2).

The invention relates to a preparation method for a duck viral hepatitis refine yolk antibody, which comprises the following steps: (1) preparation of virus seed for production; (2) preparation of venom for preparing antigen; (3) preparation of oil-emulsion inactivated vaccine; (4) preparation of eggs from hype-immunized chickens; (5) separation of yolk; (6) inactivation I and extraction; (7) inactivation II, collation and deep filtration; (8) inactivation III and degerming filtration; and (9) mixing and package, wherein the collected sterile filtered liquid is subject to liver neutralizing antibody titer determination; the antibody is diluted until the neutralizing antibody titer of the antibody is 1:256-1:1,024 by using 0.015mol / L of sterilized PBS with the pH of 7.2; 0.02 volume percent of tween-80 as a stabilizer is added in the antibody, and the mixture is evenly mixed and sub-packaged; and the obtained product is sterile and quantitative sub-packaged. The product has the advantages of high purity, strong specificity, easy preservation, no residue and the like, and has extremely good effect of preventing and treating the duck viral hepatitis.

The invention provides a feed, comprising a Chinese herbal medicine composition used for treating duck virus hepatitis and a feed carrier, wherein the Chinese herbal medicine composition further comprises oriental wormwood, uncaria, indian epimeredi herb, schizonepeta, gastrodia elata, safflower, citron, llicis routundae cortex, sculellaria barbata, common reed rhizome, pipewort, fructus forsythiae, folium apocyni veneti, radix angelicae pubescentis, codonopsis pilosula and Chinese yam. The feed containing the Chinese herbal medicine composition can overcome various defects of traditional Chinese medicine and Western medicine in the prior art, has obvious treatment effect on ducks suffering from the duck virus hepatitis, treats both principal and secondary aspects of the duck virus hepatitis, strengthens the body resistance to eliminate pathogenic factors and has the advantages of high cure rate, no toxic or side effect, low recurrence rate, no phytotoxicity and low cost.

The invention belongs to the technical field of biological detection and specifically relates to a duck viral hepatitis type 1 RT-LAMP detection kit and a detection method thereof. The invention discloses the duck viral hepatitis type 1 RT-LAMP detection kit and is characterized in that the kit contains two pairs of primers, the nucleotide sequence of which is as shown in SEQIDNO.1-4. The detection kit and its detection method provided by the invention have advantages of high speed, high sensitivity, high specificity, low cost and simple operation, can make up for the deficiency of present duck viral hepatitis detection methods, can satisfy the detection requirement of the disease at present, are easy for popularization and application at large scale, can be used to decrease the prevalence of the disease in ducks and other animals, and have a wide market prospect and great economic benefits.

The invention discloses a VP (viral protein)0 recombinant protein of DHAV (duck hepatitis A virus)-1 as well as a preparation method and an application of the VP0 recombinant protein and belongs to the technical field of bio-medicine. The preparation method of the VP0 recombinant protein comprises the following steps: a), target fragment cloning of a whole VP0 gene and expression vector construction; b), expression of the VP0 recombinant protein; c), purification of the VP0 recombinant protein. The invention further provides research on the function of the VP0 recombinant protein in detection and immune protection. A novel way is provided for detection, prevention and treatment of duck viral hepatitis, an indirect ELISA (enzyme linked immunosorbent assay) method based on the VP0 recombinant protein is established, and the method has the characteristics of good repeatability and high specificity.

The invention discloses a functional feed for treating duck virus hepatitis and a preparation method of the functional feed for treating the duck virus hepatitis. The feed comprises the following raw materials: corns, bran, soybean meal, mixed powder, stone powder, shell powder, table salt, sorghum, fish meal, crushed rice, meat meal, vegetable cakes, a vitamin additive and a mineral substance additive. The preparation method of the feed for treating the duck virus hepatitis comprises the following steps: a crushing step; a material preparation step; a granulation step; and a drying step. The functional feed has the beneficial effects of containing a traditional Chinese medicine additive with the effects of clearing heat, detoxifying, and activating and cooling blood so that the feed has the effect of strengthening body resistance to eliminate pathogenic factors and has the advantages of relatively good curative effect, no medicine residues, low cost and the like; the traditional Chinese medicine additive and the other feed raw materials are matched and are interacted so that the duck virus hepatitis can be treated and the nutritional requirements required by young ducks also can be met; and the functional feed is especially suitable for providing the nutritional requirements for the young ducks with the virus hepatitis, and a certain health effect on an organism of the consumer can be realized after a consumer eats duck meat.

The invention discloses divalent egg yolk antibodies of type I and novel duck hepatitis as well as a preparation method and an application thereof. The preparation method comprises the steps of high-immunity egg collection, high-immunity egg sterilization, fat removal with sodium alginate, precipitation with ammonium sulfate, ultrafiltration membrane concentration and purification of the antibodies, bacteria removal by inactivation and the like. The research results show that the divalent egg yolk antibodies of type I and novel duck hepatitis, which are obtained through grease removal by adopting sodium alginate and precipitation by adopting ammonium sulfate, have high purity, high titers, small side effects and obvious treatment effects and can be used for simultaneously controlling type I and novel duck hepatitis viruses safely and conveniently.

A veterinary medicine for treating the disease in digestive tract of animal and fowl is prepared from acetylmethaquine, anisodamine hydrochloride, sodium salicylate, and 4 Chinese-medicinal materials including pink plumepoppy herb, coptis root, etc.

The invention relates to a traditional Chinese herbal medicine powder for preventing and treating duck virus hepatitis, which is characterized by consisting of the following drugs according to parts by weight: 1-3 parts of artemisia capillaries, 1-4 parts of rhubarb, 1-3 parts of gardenia, 1-3 parts of gentian and 1-3 parts of radix isatidis; and the preparation method thereof comprises the following steps: weighting the drugs according to parts by weight, grinding, passing through a 200-400 mesh sieve, and then evenly mixing, thus obtaining the traditional Chinese herbal medicine powder. Thetraditional Chinese herbal medicine powder for preventing and treating duck virus hepatitis is characterized by quick effect, indeed efficacy, no drug residues or toxic side effects by adopting pure traditional Chinese herbal medicine raw materials.

The invention discloses a medicament for treating duckling viral hepatitis which comprises the following raw materials (by weight portion): radix isatidis 20-25, dyers woad leaf 17-22, gentian root 12-17, alkanna tinctoria 14-19, coptis root 10-15 and licorice root 17-22. The process for preparation consists of mixing the raw material herbs, immersing 6-9 hours in water, grilling 1.5-2 hours, obtaining filtrate, charging water, grilling 2 hours, filtrating and deslagging, finally mixing the two filtrates homogeneously.

The invention discloses a traditional Chinese medicine for treating the duck viral hepatitis. The traditional Chinese medicine is prepared from the following raw materials in parts by weight: 15-45 parts of radix isatidis, 30-90 parts of herba artemisiae, 10-30 parts of gardenia, 30-90 parts of stringy stonecrop, 15-24 parts of giant knotweed, 9-15 parts of salvia miltiorrhiza, 9-21 parts of radix paeoniae alba, 9-21 parts of radix bupleuri, 9-24 parts of turmeric, and 5-15 parts of ganoderma lucidum. Compared with the prior art, the novel traditional Chinese medicine and the preparation method provided by the invention can be used for treating and controlling ducklings with duck viral hepatitis with the remarkable treatment effect, and has the advantages of treating both symptoms and root causes, strengthening the body resistance to eliminate pathogenic factors, adjusting the body completely, balancing yin and yang, high recovery rate, no toxic and side effects, low possibility of recurrence, no risk of damage, and low cost.

The invention belongs to the field of traditional Chinese veterinary medicines, and in particular discloses a traditional Chinese veterinary medicine oral solution for preventing and treating viral diseases of poultry. The traditional Chinese medicine is composed of a liquid medicine A and a liquid medicine B, wherein the active ingredients of the liquid medicine A are composed of astragalus mongholicus, semen raphani, white paeony root, sanguisorba officinalis, herba epimedii, radix bupleuri, fructus aurantii, and liquorice; the active ingredients of the liquid medicine B are composed of coptis chinensis, rheum officinale, polygonum cuspidatum, cyrtomium fortunei, honeysuckle, fructus forsythiae, and houttuynia cordata. The traditional Chinese veterinary medicine oral solution disclosed by the invention is mainly used for treating the viral diseases of poultry and safe in use, avoids western medicine residues to guarantee the security of meat and eggs, has an obvious and durable curative effect, is fast to be absorbed, works rapidly, has strong effects, is simple to take, has sufficient medicine sources, is proper in price, and can be used for treating the viral diseases of poultry fundamentally and preventing and treating the viral diseases, such as Newcastle disease and duck virus hepatitis, of poultry.

The invention provides a monoclonal antibody against duck hepatitis A virus type A. According to the invention, mixed polypeptide is coupled with KLH carrier protein through cysteine residues so as to obtain an antigen which is used to immunize a mouse; spleen cells of the mouse are fused with SP2 / 0 bone marrow cells, and screening is carried out so as to obtain a hybridoma cell line 3L1-6 with a microbial accession number of CCTCC No: C2013134. The monoclonal antibody prepared from the hybridoma cell line has strong specificity and good stability and can be extensively used in preparation of detection reagents for antibodies against duck hepatitis A virus type A. An ELISA kit developed from the antibody has strong specificity and good stability and can be used for detection of antibodies against duck hepatitis A virus type A.

The invention relates to the field of animal medicine, in particular to a method for detecting duck plague virus (DPV) antibodies. The method specifically comprises the steps of preparation of solid phase antigens, primary antibody combination, secondary antibody combination, color development, detection, judgment and the like. The method is a purified recombinant UL55 protein-based indirect enzyme-linked immuno sorbent assay (ELISA) method, and has good specificity; the results of detecting the positive serums of duck virus hepatitis viruses (DHV), riemerella anatipestifer (RA), duck escherichia coli (E.coli), duck-derived salmonella, duck swollen head hemorrhagic disease viruses and duck-derived influenza viruses are negative; and the repeated tests in enzyme-labeled plates or between enzyme-labeled plates display that variation coefficients are less than 10 percent, and the method can detect out the positive serums of DPV weak virus vaccine immune ducks, wherein the DPV weak virus vaccine are diluted according to a ratio of 1:6400.

The invention relates to the technical field of animal virology and animal infectious disease, and particularly relates to duck virus hepatitis suicide DNA vaccine, and the building method and the application thereof. The duck virus hepatitis suicide DNA vaccine provided by the utility model comprises DHV-VPI gene and alphavirus copying sub carrier pSCA 1, the constructing of the DNA vaccine comprises the following steps: the total RNA of DHV virus is extracted and purified, specific primers are designed, through the augmentation of RT-PCR, the total length cDNA gene order of DHV-VPI can be cloned, and finally, a purified VP 1 total length Cdna segment is restructured into the eukaryotic expression plasmid of alphavirus copying sub carrier pSCA 1. Compared with the prior art, the required immunizing dose of the vaccine provided by the invention is small, and the vaccine is safe and efficient; the vaccine can be eliminated by an organism with the Apoptosis of cells. Therefore, DNA plasmid are prevented from being possibly integrated into host cell chromosomes or causing hidden troubles such as immune tolerance and the like, and the duck virus hepatitis suicide DNA vaccine is efficient and stable, and the operation is convenient and quick.

The invention relates to a heat-resisting protective agent for duck virus hepatitis live vaccines and a preparation method and application of the protective agent. The method comprises the following steps of: mixing and preparing a freeze-dried protective agent matrix in a certain ratio, performing sterile treatment on the protective agent, dissolving the protective agent components for autoclaved sterilization into double distilled water, and sterilizing for 15 to 30 minutes at the temperature of between 108 and 121 DEG C; and dissolving the protective agent components without autoclaved sterilization resistance into double distilled water in a ratio, removing bacteria by using a 0.22mu m filter membrane, mixing the both to form the heat-resisting protective agent, mixing the heat-resisting protective agent and duck virus hepatitis liquid in a ratio of 1: (1-1.2), and performing freeze-drying by adopting a corresponding freeze-drying curve. Compared with the prior art, a good resource is provided for preparation of the duck virus hepatitis live vaccines, the backward status of a domestic vaccine preservation technology is overcome, the live vaccines are preserved for 24 months at the temperature of between 2 and 8 DEG C, and the titers of the vaccines on each feather are kept to be more than or equal to 106.0EID50.