93 results about "Duck hepatitis virus" patented technology

The invention discloses a Korean novel duck hepatitis viral antibody ELISA (Enzyme-Linked Immunosorbent Assay) detection kit. The detection kit contains an ELISA board coated by Korean novel duck hepatitis VP1 (Phenotypic Variance1) recombination protein, a sample diluent, concentrated washing liquid, an enzyme conjugate working solution, a chromogenic reagent (A), a chromogenic reagent (B), a stopping solution, a positive contrast solution and a negative contrast solution. The VP1 recombination protein is obtained by using the following method: using Korean novel duck hepatitis viruses as a material, augmenting and cloning the VP1 gene through an RT-PCR (Reverse Transcriptase-Polymerase Chain Reaction) method to obtain recombinant expression plasmid pMD (physical medium dependent)-VP1; then, directionally inserting to an expression vector pET-32a (+) and screening to obtain recombinant expression plasmid pET-32a(+)-VP1; and inducing, expressing and purifying by ITPG (Isopropyl beta-D-Thiogalactopyranoside) to obtain VP1 recombination protein. The detection kit is used for detecting the Korean novel duck hepatitis and has strong specificity, high sensitivity, simplicity of operation, easiness of popularization and application in a large-area range and wide market prospects.

The invention discloses a molecular biological identification method for I type duck hepatitis virus, which comprises the steps of treating samples to be tested, preparing RNA of the samples, obtaining cDNA of the samples by reverse transcription, identifying I type duck hepatitis virus by the fluorescent quantitive RT-PCR method or the nested PCR method, etc. The molecular biological identification method has the advantages of fast diagnosis, high sensitivity and wide applied range, can be used for identifying DHV-1 from the tissue samples, cotton swabs, ducks or chicken embryo allantoic liquid or cell cultures.

The invention discloses a duck hepatitis virus type I LAMP (loop-mediated isothermal amplification) detection kit. A LAMP technology is adopted; and two specific inner primers, two specific outer primers and two specific ring primers are designed according to a gene conserved region shared by duck hepatitis viruses type I. Due to the application of the duck hepatitis virus type I LAMP detection kit and a detection method established by the invention, the defects of long detection time, large workload, cross pollution, complex operation and the like in the prior art are overcome. The duck hepatitis virus type I LAMP detection kit has the strong specificity and high sensitivity, is rapid, has low cost and simpler operation method and is particularly suitable for the requirement on field rapid detection of the duck hepatitis virus type I in the primary-level areas.

The invention discloses a PCR-RFLP primer for distinguishing DHV-1 and a new serotype and a method for distinguishing the DHV-1 and the new serotype. The primer is an upstream primer P1:5'-CAATTGAGGACATGGCTAAGAAA-3' and a downstream primer P2:5'-TCAGAGASCCRTCRAAWCCAGAAA-3. According to the method, distinguishing is conducted according to the DHV-1 and the restriction enzyme cutting site difference contained in a new serotype coding area and includes the steps that RNA extraction is conducted, and after corresponding target fragments are obtained through RT-PCR amplification, RFLP analysis is conducted after XhoI restriction enzyme cutting. The distinguishing method is simple and fast in operation and high in accuracy.

The invention discloses an indirect ELISA method and kit for detecting serum 3-type duck hepatitis virus a antibody and belongs to the technical field of serum antibody detection. The indirect ELISA method includes that serum 3-type duck hepatitis virus VP1 protein is utilized as an envelope antigen with the peridium quantity as 0.1-1mug per hole, HRP-mice anti-duck IgY is utilized as the HRP with the use concentration as 0.2-2mug/mL, and the coloration time is 10 minutes. The kit comprises a VP1 protein antigen envelope board, the HRP-mice anti-duck IgY, sample diluent, a scrubbing solution, TMB solutions A and B and a stop solution. The method and the kit can be used for detecting the serum 3-type duck hepatitis virus a antibody in duck serum and duck egg yolk, and the serum does not have cross reaction with positive serum of other viruses. Compared with a traditional neutral reaction detection method, the method has the advantage of being convenient and accurate.

The invention provides a muscovy duck hepatitis virus and a method for preparing a muscovy duck hepatitis virus refined egg yolk antibody by adopting the muscovy duck hepatitis virus. The muscovy duck hepatitis virus strain GS14 is preserved in the China Center for Type Culture Collection on April 27, 2016, and the accession number is CCTCC No: V201623. The method for preparing the muscovy duck hepatitis virus refined egg yolk antibody by adopting the muscovy duck hepatitis virus comprises the following steps: preparing antigens by adopting the muscovy duck hepatitis virus strain GS14; carrying out inactivation on the antigens; preparing antigens for vaccines; preparing inactivated vaccines; and preparing the muscovy duck hepatitis virus refined egg yolk antibody. The muscovy duck hepatitis virus and the preparation method have the beneficial effects that the prepared muscovy duck hepatitis virus refined egg yolk antibody can prevent and treat the white liver disease of the muscovy duck, so that an effective means is provided for preventing and treating the white liver disease of the muscovy duck clinically.

The invention provides preparation methods for duck hepatitis virus immunogen and hyperimmune serum and application of the duck hepatitis virus hyperimmune serum. According to the preparation methods and the application of the duck hepatitis virus hyperimmune serum, the duck hepatitis virus immunogen is obtained through inoculating a serum 1 type duck hepatitis virus CH60 strain DHAV-1 (Duck Hepatitis A Virus type 1) or a serum 3 type duck hepatitis virus CH1 strain DHAV-3 (Duck Hepatitis A Virus type 3) to an allantoic cavity of a chick embryo of 9-10 days old or a duck embryo of 10-12 days old and carrying out proliferation and treatment, and the hyperimmune serum is obtained through mixing the duck hepatitis virus immunogen with a Freund's complete adjuvant or Freund's incomplete adjuvant to prepare solutions of different concentrations, carrying out repeated immunization on immune animals and then sampling and collecting blood and can be applied to the diagnosis and detection on a duck hepatitis virus. The preparation methods provided by the invention have the advantages that the conditional requirements are low, the operation is simple, and the obtained immunogen can meet the requirements on the preparation of specific antiserum.

The invention discloses a Riemerella anatipestifer OmpA/MotB truncated recombinant protein, its antibody and a preparation method and an application thereof. The recombinant protein has immunoreactivity similar to natural OmpA/MotB protein, can bind to RA antibody specifically, will not generate cross reactions with Salmonella anatum positive serum, duck colibacilosis positive serum, duck swollen head septicemia virus positive serum, avian influenza virus (H5) positive serum, duck plague virus positive serum, duck hepatitis virus positive serum and the like, and can be used as a detection antigen for detecting whole bacteria antibody. As the recombinant protein is not whole bacteria, there is no risk of spreading bacteria when the recombinant protein is applied to detection. In addition, the preparation method of the recombinant protein is simple, is suitable for large-scale industrial production, can realize standardization of products and is beneficial to result comparison between different laboratories.

The invention relates to a novel freeze-drying protective additive for duck virus hepatitis live vaccines. The novel freeze-drying protective additive has the advantages that the vaccine is low in loss of antigen activity during freeze drying, and the problem that the duck hepatitis virus is sensitive to the freeze drying is effectively solved; the duck virus hepatitis vaccines freeze-dried by the protective additive are good in heat resistance, and storage life thereof is longer than 24 months at the temperature of 2-8 DEG C, good activity is still maintained in ten days after a resistance aging test at the temperature of 37 DEG C, so that the problem of a strict low-temperature cold chain of the vaccines upon storage and transportation is solved, and meanwhile vaccine storage and transportation costs are greatly lowered.

The invention discloses a method for establishing goose embryo epithelial cell line and the established goose embryo epithelial cell line. The invention relates to the method for establishing the goose embryo epithelial cell line, which is characterized in that a primary goose tissue adherent method, a differential velocity enzymatic digestion and a monoclonal screening method are combined, so that primary culture condition can be optimized. The method has the advantages of simple operation process, and convenient popularization and application. The invention also relates to the goose embryo epithelial cell line established by the method, a preservation number of the goose embryo epithelial cell line is CCTCC NO: C2014137, The establishment of the goose embryo epithelial cell line solves the problem of no well-established goose source cell line in prior art. The invention also relates to a kit used for culturing and/or proliferating goose parvovirus, muscovy duck parvovirus and type I duck hepatitis virus and novel duck hepatitis virus, and is characterized in that a host cell to be infected is/or comprises the goose embryo epithelial cell line. The method for establishing the goose embryo epithelial cell line verifies the sensitive characteristic of the goose parvovirus, muscovy duck parvovirus and type I duck hepatitis virus and novel duck hepatitis virus.

The invention belongs to the bird immunology technology area, it concretely relates to a kit of acceptable for the one type duck hepatitis virus antibody emulsion agglutination and the application of testing one type duck hepatitis virus antibody emulsion agglutination. The kit of this invention contains box body and core reagent in the box body, positive check sample, negative check sample, sample cell, scale sample sucker and the carrier of observed result. The characteristic is that core reagent is latex antigen from sublimating e duck hepatitis virus AV2111 and inactivating one type duck hepatitis virus sensitization polystyrene latex, the positive check sample is one type duck hepatitis virus antiserum, the negative check sample is the green duck of a day old of no one type duck hepatitis mother source antibody. The carrier is the paper characteristic latex agglutination card of having hollowness. The invention also discloses the preparation method and application of this kit. The kit has the significance merit of high specificity, high sensitivity, handy operation, and quickly diagnose.

The invention provides a GAOH derivative of a new structure. The compounds have great curative effects on inflammatory bowel diseases, hepatitis viruses, foot swelling, ear swelling, arthritis, pneumonia, nephritis, rhinitis, cerebral apoplexy, myocardial ischemia, atherosclerosis, arrhythmia, autoimmune disease, senile dementia, depression, schizophrenia, malignancy and tumor adjuvant therapy, virus infectious diseases, peptic ulcer, analgesia, antianaphylaxis, antiendotoxin, antishock and diabetes or diabetic complication.

The invention discloses a triple inactivated vaccine for duck circovirus disease, novel duck reovirus disease and duck virus hepatitis and a preparation method of the triple inactivated vaccine. The triple inactivated vaccine comprises an effective amount of inactivated antigens and immunologic adjuvants in immunization, and the inactivated antigens include a duck circovirus antigen, a novel duck reovirus antigen and a duck hepatitis virus antigen. The invention further discloses a method for preparing the triple inactivated vaccine. The triple inactivated vaccine for the duck circovirus disease, the novel duck reovirus disease and the duck virus hepatitis is good in safety and does not generate mutual interference or influence of antigen components. Immune protection efficacy tests prove that the triple inactivated vaccine can prevent duck reovirus, duck circovirus and duck hepatitis virus at the same time, can avoid adverse reactions caused by multiple inoculation immunization, and has the advantages of simple preparation method, convenience, multiple effects, low cost, high vaccine titer content and the like.

The invention provides a one-step process PCR detection method for duck hepatitis viruses and duck plague viruses. The method comprises the following steps: respectively selecting highly conservative genes in the duck hepatitis viruses and the duck plague viruses as a target fragment I and a target fragment II, and designing and synthesizing corresponding specific primers I and II; respectively extracting genome RNAs and DNAs of the duck hepatitis viruses and the duck plague viruses, mixing the genome RNAs and DNAs as templates, and carrying out double PCR amplification; and analyzing the obtained double PCR products by virtue of agarose gel electrophoresis. The one-step process PCR detection method provided by the invention has the advantages that the operation is simple, and the time and the labor are saved; PCR and RT-PCR can be carried out in the same reaction system, and DNA viruses (duck plague viruses) and RNA viruses (duck hepatitis viruses) can be simultaneously amplified and detected, so that the detection time is greatly shortened, and the detection human cost is greatly lowered; the specificity is good, highly specific segments can be amplified, and impurities in products are extremely few; and the one-step process PCR detection method is suitable for being generalized and used.

The invention relates to a preparation method and use of a soluble type-I duck hepatitis virus 3C protein. The preparation method comprises connecting nucleotides at 3th to 574th sites shown in the formula of SEQ ID NO. 3 to polyclonal cleavage sites of a pET-32(a)+vector, carrying out activation in an Amp-resistant LB culture medium based on escherichia coli BL21 as host bacteria, and carrying out induced expression under conditions of the IPTG final concentration of 0.2-1.4 mmol/L and a temperature of 16-37 DEG C for 2-12h to obtain a soluble type-I duck hepatitis virus 3C protein. The protein is obtained by soluble expression, is natural, is free of denaturant-based denaturation and renaturation and can be used for 3C protease activity detection and 3C protease inhibitor screening. Thepreparation method has a great importance for research, development and screening of a drug for resisting type-I duck hepatitis viruses.