22 results about "Duck hepatitis A virus" patented technology

The invention relates to the field of pathogen detection, in particular to a primer probe group, a gene chip and a kit for detecting seven waterfowl infectious disease pathogens and application. The primer probe group provided by the invention can be used for simultaneously detecting the goose parvovirus, the duck viral enteritis virus, the muscovy duck parvovirus, the duck hepatitis A virus type 1, the duck hepatitis A virus type 3, the duck tembusu virus and the novel duck reovirus, and is strong in specificity and high in sensitivity. Furthermore, a gene chip prepared by using the primer probe group provided by the invention not only has strong specificity and high sensitivity (1.0 * 100 copies / [mu] L), but also has good repeatability and stability, can be stably stored within at least 180 days, has sensitivity superior to that of a standard fluorescent quantitative PCR method, can rapidly and efficiently detect main epidemic disease pathogens of waterfowl together, and has good application prospects. The method can be used for epidemiological investigation and disease diagnosis of waterfowls.

The invention discloses an NDRV [sigma] C circular RNA vaccine and application thereof, and belongs to the technical field of vaccines.Duck hepatitis A virus type 1 (DHAV-1) UTR and T4 bacteriophage I type intron self-splicing cyclization systems are adopted to construct circular RNA (CirRNA-[sigma] C) for expressing NDRV [sigma] C protein, and a vaccine preparation is prepared through a chitosan-polyethyleneimine (CS-PEI) nano delivery system. The feasibility of a circular RNA vaccine system based on DHAV-1UTR in poultry vaccines is confirmed for the first time, the established nano delivery technology and circular RNA vaccine platform have the characteristics of high stability, good safety, strong immunogenicity and the like, and an important technical path is provided for development of novel vaccines of NDRV and other viral epidemic diseases.

The invention discloses a self-assembled ferritin nano antigen particle and application thereof in preparation of a duck hepatitis A vaccine. The VP1 part in the duck hepatitis A virus P1 protein is connected with the N end of a self-assembled ferritin nanoparticle subunit through a connecting sequence to obtain the fusion protein, and the amino acid sequence of the fusion protein is shown as SEQ ID NO.1; in order to improve the expression quantity or expression efficiency of the fusion protein in bombyx mori, a nucleotide sequence of a coding gene of the fusion protein is optimized and modified according to the codon preference of bombyx mori, and an optimized gene sequence with the nucleotide sequence shown as SEQ ID NO.2 is obtained; the fusion protein is subjected to single-site or multi-site mutation, the mutant with obviously improved titer is obtained through screening, and the soluble expression quantity of the mutant is obviously improved. According to the invention, a silkworm or insect cell eukaryotic expression system is used for expressing recombinant protein, and an expression product causes a neutralizing anti-duck hepatitis A virus antibody.

The invention discloses a method for establishing duck hepatitis virus antibody ELISA (enzyme-linked immuno sorbent assay) detection, which comprises the following steps: firstly, preparing experimental materials, and then cloning and antigenicity analysis of structural genes, inducible expression of positive recombinants in escherichia coli, expression and purification of recombinant proteins, antigenicity analysis of the recombinant proteins and establishment of a recombinant antigen VP3 indirect ELISA method. Finally, VP0, VP3 and VP1 recombinant proteins can react with duck anti-DHV-1 positive serum and have good antigenicity, indirect ELISA results show that under the same coating concentration, the P / N value of the fusion protein GST-VP3 serving as a coating antigen is higher than that of the fusion protein GST-VP3 serving as the coating antigen and that of the fusion protein GST-VP3 serving as the coating antigen, it is indicated that the antigenicity of the fusion protein GST-VP3 is better than that of the fusion protein GST-VP3 serving as the coating antigen and that of the fusion protein GST-VP3 serving as the coating antigen and that of the fusion protein GST-VP3 serving as the coating antigen and that of the fusion protein GST-VP3 serving as the coating antigen and that of the fusion protein and the result is consistent with the prediction result.

The invention provides a triple tetravalent inactivated vaccine for preventing and treating duck viral diseases and application of the triple tetravalent inactivated vaccine, and belongs to the technical field of biological vaccines. A water phase of the vaccine is prepared by matching a type 1 duck hepatitis virus antigen solution, a type 3 duck hepatitis virus antigen solution and a duck reovirus antigen solution, and then adding a duck parvovirus VP2 protein solution to obtain a mixed antigen solution with the final concentration of 40 mu g / ml, the virus content in the type 1 duck hepatitis virus antigen solution and the virus content in the type 3 duck hepatitis virus antigen solution are 106.5 ELD50 / 0.2 ml, and the virus content in the duck reovirus antigen solution is 106.5 ELD50 / 0.2 ml. And the virus content of the duck reovirus antigen is 105.5 TCID50 / ml. The four antigen components in the vaccine are free of mutual interference, good in safety and exact in immune effect, the vaccine can achieve the purpose of preventing multiple diseases by one injection, the stress response of the duck immune vaccine is reduced, meanwhile, the four virus diseases are susceptible to small-day-age ducks, and invasion of the four viruses can be prevented at the same time after immunization. The vaccine preparation method is simple and convenient, and the production cost is greatly reduced.

The present invention relates to the field of pathogen detection, in particular to a primer-probe set, gene chip, kit and application for detecting seven kinds of waterfowl infectious disease pathogens. The primer-probe set provided by the present invention can simultaneously detect goose parvovirus, duck viral enteritis virus, Muscovy duck parvovirus, duck hepatitis A virus type 1, duck hepatitis A virus type 3, duck Tembusu virus and novel duck reovirus, and has strong specificity and high sensitivity. Further, the gene chip prepared by using the primer-probe set provided by the present invention not only has strong specificity and high sensitivity (1.0×10<supgt;0< / supgt> copies / μL), but also has good repeatability and stability, can be stably stored for at least 180 days, and its sensitivity is also better than that of the standard fluorescence quantitative PCR method. It can quickly and efficiently co-detect the above-mentioned main pathogens of waterfowl diseases and can be used for the epidemiological investigation and disease diagnosis of waterfowl.

This invention discloses self-assembled ferritin nanoparticles and their application in the preparation of duck hepatitis A vaccine. The invention connects the VP1 portion of the duck hepatitis A virus P1 protein to the N-terminus of a self-assembled ferritin nanoparticle subunit via a linker sequence to obtain a fusion protein, the amino acid sequence of which is shown in SEQ ID NO.1. To improve the expression level or efficiency of the fusion protein in silkworms, the nucleotide sequence of the encoding gene of the fusion protein is optimized and modified according to the codon preference of silkworms, resulting in the optimized gene sequence shown in SEQ ID NO.2. The invention also involves single-site or multi-site mutations of the fusion protein, screening for mutants with significantly improved titers and significantly increased soluble expression levels. This invention utilizes a silkworm or insect cell eukaryotic expression system to express the recombinant protein, and the expression product induces neutralizing anti-duck hepatitis A virus antibodies.

The invention discloses a recombinant duck enteritis virus for expressing 3-type duck hepatitis A virus immunogenic genes and a construction method and application thereof. The recombinant duck enteritis virus is characterized in that an eukaryotic expression cassette containing a type 3 duck hepatitis A virus immunogenic gene is inserted into a non-coding region between UL26 and USL27 genes of a duck enteritis virus genome. According to the invention, a duck hepatitis A virus (DHAV) immunogenic gene is inserted into a duck enteritis virus (DEV) genome at a fixed point by virtue of a CRISPR / Cas9 gene editing technology, so that the recombinant duck enteritis virus for expressing the duck hepatitis A virus type 3 immunogenic gene is constructed and obtained. Moreover, based on the recombinant duck enteritis virus for expressing the 3-type duck hepatitis A virus immunogenicity gene constructed by the invention, the exogenous DHAV immunogenicity gene can be stably expressed in the recombinant virus, and the recombinant virus has good in-vitro replication ability and hereditary stability and can be used for preparing animal vaccines.

The application discloses application of coenzyme Q10 and a composition containing coenzyme Q10 in preparation of a medicine for preventing and treating duck hepatitis A virus type 1, and relates to the technical field of veterinary antiviral medicines. The application finds through experiments that coenzyme Q10 and the composition containing coenzyme Q10 have a protective effect on duck hepatitis A virus type 1 infection. The composition contains coenzyme Q10 and egg yolk antibodies. The combination of coenzyme Q10 and egg yolk antibodies can relieve inflammatory reactions caused by duck hepatitis A virus type 1, reduce liver oxidative application, alleviate liver function damage caused by duck hepatitis A virus type 1, and reduce the viral load of duck hepatitis A virus type 1, so that the dual synergy of 'antiviral' and 'anti-inflammatory liver protection' is realized. The application first finds the application value of the combination of coenzyme Q10 and egg yolk antibodies in the prevention and treatment of DHAV-1 infection, and provides a new technical strategy for the prevention and treatment of duck viral hepatitis.

The application discloses a duck hepatitis virus 1 type DHAV-1 / HNSQ and application thereof, and belongs to the field of animal virology and genetic engineering. The duck hepatitis virus 1 type DHAV-1 / HNSQ has been preserved in the China Center for Type Culture Collection on March 5, 2026, and the preservation number is CCTCC NO:V202622. The VP1 gene of the variant strain has a deletion of three consecutive nucleotides. Animal experiments show that, after the variant strain infects ducklings, in addition to causing typical duck viral hepatitis lesions, the variant strain can also stably cause a new phenotype, such as thinning of the heart wall, reduction of myocardial toughness and liquefaction necrosis of pancreatic tissue, which has never been reported in the prior art, thereby providing important biological materials for effective prevention and control of duck hepatitis and pathogenic mechanism research.

The application discloses a duck hepatitis A virus type 3 VP0 polypeptide and a preparation method and application of a polyclonal antibody thereof, and belongs to the technical field of biochemistry and molecular immunology. The duck hepatitis A virus type 3 VP0 polypeptide has an amino acid sequence as shown in SEQ ID NO. 2. A polyclonal antibody is prepared by immunizing Kunming mice with the polypeptide. The polyclonal antibody of the duck hepatitis A virus type 3 VP0 polypeptide is obtained by immunizing an antiserum with a VP0 polypeptide coupled with KLH as an antigen. The polyclonal antibody preparation method is simple, low in cost, high in antibody titer, and capable of specifically recognizing the duck hepatitis A virus type 3 VP0 protein in eukaryotic cells.

The invention belongs to the technical field of virus detection, and particularly relates to a PCR (Polymerase Chain Reaction) detection kit for four vertical transmission viruses of waterfowl and application of the PCR detection kit. The kit can rapidly, specifically and simultaneously amplify and diagnose duck hepatitis virus type I (DHV-1), duck Tembusu virus DTMUV, Muscovy duck reovirus MDRV and duck astrovirus DAstV, has high specificity and sensitivity, can accurately and rapidly identify DHV-1, DTMUV, MDRV and DAstV, can complete fluorescent quantitative PCR detection in about 50 minutes, and is simple to operate and practical. The detection period is shortened to 1.5 hours from 6-8 hours of a traditional method, the sensitivity is improved by 10-1000 times, the early diagnosis capability of epidemic diseases is remarkably improved, and the epidemic disease misjudgment rate of a farm is obviously reduced. Abuse of vaccines and drugs is reduced, and sustainable development of the poultry breeding industry is promoted.

The invention belongs to the technical field of biology, and particularly relates to a recombinant baculovirus capable of simultaneously expressing gene type 1 and gene type 3 duck hepatitis A virus VP1 protein and application of the recombinant baculovirus in preparation of a duck hepatitis A divalent subunit vaccine. The recombinant baculovirus co-expressing the gene type 1 and gene type 3 duck hepatitis A virus VP1 protein is constructed, the virus titer is high, and the two proteins can be expressed. Therefore, the recombinant baculovirus provided by the invention can be used for research and development and epidemic disease prevention and control of novel vaccines for duck viral hepatitis A.

The invention belongs to the technical field of biology, and particularly relates to an annular KANSL1 gene and application thereof in regulation and control of replication of type 3 duck hepatitis A virus. The overexpression of the annular KANSL1 gene can significantly promote the replication of the intracellular type 3 duck hepatitis A virus, and the replication of the intracellular type 3 duck hepatitis A virus can be significantly inhibited by inhibiting the expression of the annular KANSL1 gene through small interfering RNA.

The invention provides application of a duck APOBEC1 gene and a related biological material thereof in resisting duck hepatitis virus, and particularly belongs to the technical field of biology. Transcriptome sequencing analysis is carried out on livers of DHAV-3 resistant ducks and susceptible ducks, a candidate gene APOBEC1 responding to resistance of Beijing ducks is screened out, and the invention provides application of APOBEC1 protein and related biological materials to preparation of drugs for inhibiting DHAV-3 infection and / or preparation of inhibitors for inhibiting DHAV-3 virus. The antiviral function of the gene is identified through overexpression and gene transcription level inhibition, a foundation is laid for deep research on the function and disease resistance mechanism of APOBEC1, and candidate gene resources are provided for disease resistance breeding of Beijing ducks.

The present invention relates to the field of viral molecular biology detection by establishing a dual real-time fluorescence quantitative PCR primer set and kit for duck hepatitis A virus type 1 and duck astrovirus type 3. This method can rapidly detect duck hepatitis A virus type 1 and duck astrovirus type 3. The method is simple and rapid, with a single sample detection time of 90 minutes and a sensitivity of up to 10 copies / μl. Simultaneous detection is possible for larger sample sizes, and the method exhibits excellent sensitivity, repeatability, specificity, and stability, enabling rapid and accurate detection of duck hepatitis A virus type 1 and duck astrovirus type 3.

The present invention belongs to the field of biological medicine, and particularly relates to a duck cardiomyocyte line and its construction method and application. The present invention provides a duck cardiomyocyte line named Sheldrake Duck Myocardial Cells SDMC, which was deposited on July 5, 2024 at the China Center for Type Culture Collection, Wuhan University, Wuhan, China, with the deposit number of CCTCC NO: C2024190. It has been passaged to 80 generations at present, and the morphology is mainly spindle-shaped cells; the virus sensitivity test shows that this duck cardiomyocyte line is sensitive to infectious bursal disease virus, avian adenovirus and Duck Hepatitis A Virus Type I; this cell line can be used for culturing, isolating and detecting avian viruses, preparing avian vaccines, and screening drugs for preventing or treating diseases caused by avian virus infections.

The invention discloses a 1-type duck hepatitis A virus BJ80 strain, a vaccine and application of the 1-type duck hepatitis A virus BJ80 strain and the vaccine. The preservation number of the duck hepatitis A virus type 1 BJ80 strain is CCTCC (China Center For Type Culture Collection) NO: V202567. The 1-type duck hepatitis A virus attenuated strain with the preservation number of CCTCC NO: V202567 is obtained through chicken embryo passage and attenuation, and the technical problems that an existing DHAV-1 virulent strain is high in virulence and poor in safety, and an existing vaccine is strong in virulence, unstable in immune effect, short in storage life and the like are successfully solved; the low virulent strain is safe to ducklings, high in hereditary stability, free of strong reversion after continuous passage and good in immunogenicity, can induce generation of effective immune protection after inoculation, and provides a reliable core seed virus for preparation of safe and efficient bivalent live vaccines for duck virus hepatitis.

The invention discloses a construction method of a rapid virus detection system, a rapid 1-type duck hepatitis A virus detection system, a detection method and a detection kit, and relates to the technical field of rapid pathogen detection. The construction method of the rapid detection system for the type 1 duck hepatitis A virus comprises the following steps: step 1, primary screening; 2, designing a primer; step 3, verification; step 4, construction of standard quality grains; and 5, optimizing the system, and adjusting the concentration of crRNA, the concentration of Cas12a and the reaction time of Cas12a by using standard plasmids as a positive detection template to obtain a rapid detection system with the reaction time not longer than 30 minutes. The invention also provides a detection method based on the detection system and an optimized detection kit, and the detection kit can realize rapid, accurate, high-specificity and high-sensitivity detection of the 1-type duck hepatitis A virus, and is beneficial to popularization and application.

The present invention provides a multiplex qPCR primer probe set, a detection system and an application for detecting duck Tembusu virus, duck reovirus, duck hepatitis A virus type 1 and duck hepatitis A virus type 3, and belongs to the technical field of virus detection. The multiplex qPCR detection primer probe set of the present invention includes an upstream primer, a downstream primer and a fluorescent probe for detecting duck Tembusu virus; an upstream primer, a downstream primer and a fluorescent probe for detecting duck reovirus; an upstream primer, a downstream primer and a fluorescent probe for detecting duck hepatitis A virus type 1, and an upstream primer, a downstream primer and a fluorescent probe for detecting duck hepatitis A virus type 3. The primer probe set of the present invention has good specificity and sensitivity, and can detect four waterfowl infectious disease pathogens at the same time.

This invention discloses the application of duck IL7 protein as an adjuvant in the preparation of duck hepatitis A virus type 3 inactivated vaccine, belonging to the field of biopharmaceuticals. The research of this invention shows that duck IL7 protein as an adjuvant can enhance the immunoprotective efficacy of the DHAV-3 inactivated vaccine and also enhance its heterologous protective ability. This provides a theoretical basis for the development of duck IL7 as a novel duck vaccine adjuvant.

The invention belongs to the technical field of biology, and provides an in-situ ELISA (enzyme-linked immuno sorbent assay) detection method for a duck hepatitis A virus type 3 egg yolk antibody. The duck hepatitis A virus type 3 strain is named as DHAV-3 / SD / 2024 / 06 strain, and the preservation number of the duck hepatitis A virus type 3 strain is CCTCC NO: V2024115. The invention establishes a duck hepatitis A virus type 3 egg yolk antibody in-situ ELISA (enzyme-linked immunosorbent assay) detection method, and makes up the blank that no similar product exists in the current market.