28 results about "Goose astrovirus" patented technology

The invention relates to the technical field of biological agents, in particular to a goose astrovirus egg yolk antibody and a preparation method and application thereof. The invention provides a goose astrovirus egg yolk antibody. The goose astrovirus egg yolk antibody is prepared by adopting an inactivated vaccine as an antigen, wherein the inactivated vaccine is prepared from a goose astrovirus GAstV / SD / 36-95P strain with the preservation number of CCTCC (China Center For Type Culture Collection) NO: V202445. The neutralizing titer of the goose astrovirus egg yolk antibody provided by the invention is high and can reach 10.5 log2, after goslings are infected by the goose astrovirus, the goslings can be protected from disease attack and death by injecting the egg yolk antibody, and the protection rate of the egg yolk antibody can reach 100%.

The invention discloses a type 2 GAstV capsid protein monoclonal antibody and an antigen epitope peptide of the type 2 GAstV capsid protein monoclonal antibody. The monoclonal antibody is prepared by immunizing a mouse with capsid protein GAstV Cap of the type 2 goose astrovirus as an antigen, and is named as 1F1; the monoclonal antibody 1F1 comprises a heavy chain variable region and a light chain variable region, and the amino acid sequences of the heavy chain variable region and the light chain variable region are sequentially shown as SEQ ID No.3 and SEQ ID No.5. In order to determine the B cell antigen epitope of the screened monoclonal antibody, the invention also constructs a GAstV Cap truncated fusion protein, and the antigen recognition epitope is determined through indirect ELISA and Western blotting tests, so that screening is carried out step by step until the shortest linear epitope recognized by the monoclonal antibody is determined, and the amino acid sequence of the shortest linear epitope is shown as SEQ ID No.10. Therefore, the monoclonal antibody and the antigen epitope of the monoclonal antibody can be applied to research on structures and functions of GAstV capsid proteins and preparation of serological diagnostic tools and epitope vaccines.

The invention relates to the field of molecular biology, in particular to a primer probe combination for detecting genotype-2 goose astrovirus and a cdPCR kit. The nucleotide sequences of the primer and the probe are shown as SEQ ID NO.1-3. The cdPCR kit comprises the primer and probe combination. The cdPCR kit disclosed by the invention is applied to detection of GAstV-2, has the advantages of high sensitivity, strong specificity and stable repeatability, can realize rapid and accurate detection of early stage of GAstV-2 infection, and provides technical support for epidemiological investigation of GAstV-2 and early prevention and control of gout of goslings.

The invention relates to the technical field of biological products, in particular to an indirect ELISA (Enzyme-Linked Immunosorbent Assay) detection kit and a detection method for a goose astrovirus antibody. The kit comprises an elisa plate, an elisa secondary antibody, goose astrovirus positive control serum, goose astrovirus negative control serum, a diluent, a washing solution, a developing solution and a stop solution, wherein a coating antigen of the ELISA plate is a goose astrovirus VP27 recombinant protein, the goose astrovirus is a goose astrovirus GAstV / SD / 36, and the goose astrovirus VP27 recombinant protein corresponds to an ORF2 region. The goose astrovirus GAstV / SD / 36 is used as a template, the virus antigen recognition site VP27 protein is selected as an antigen for preparing the indirect ELISA kit, the indirect ELISA detection kit of the goose astrovirus antibody is prepared, and the accuracy of the detection result of the kit can be improved.

The invention belongs to the field of biology, and relates to a broad-spectrum monoclonal antibody aiming at avian influenza virus M1 protein and application of the broad-spectrum monoclonal antibody, the monoclonal antibody has specific reaction with A549 cells infected by avian influenza, has no specific reaction with Newcastle disease virus, duck tembusu virus, goose astrovirus and infectious laryngitis virus, has good specificity, and can be used for preparing the broad-spectrum monoclonal antibody. The 5G9 monoclonal antibody has good reactivity with A549 cells infected by H1-H11 subtype AIV, has good broad spectrum, can be used for detecting M1 protein sub-localization after the AIV infected cells are detected by using the 5G9 monoclonal antibody, and can be used for indicating the infection process of the AIV.

The invention discloses a type 1 goose astrovirus virus-like particle and a preparation method and a pharmaceutical composition thereof, and the preparation method comprises the following steps: cloning a gene segment for coding type 1 goose astrovirus ORF2 protein to a baculovirus transfer vector, and constructing to obtain a recombinant plasmid; transforming the recombinant plasmid into a DH10Bac competent cell to obtain a recombinant baculovirus rod grain; transfecting the recombinant baculovirus stem into an insect baculovirus expression system to obtain a recombinant baculovirus; and inoculating the recombinant baculovirus into an insect baculovirus expression system, culturing, collecting a cell culture, and purifying to obtain the purified goose astrovirus type 1 virus-like particles. The GAstV-1 VLPs and the vaccine thereof provided by the invention not only solve the technical problem of research and development of the GAstV-1 vaccine, but also have the advantages of clear structure, high safety, good immunogenicity and exact protective efficacy, and have remarkable creativity and industrial application value.

The invention discloses a universal one-step RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection primer and a kit for goose astrovirus, and belongs to the field of biotechnology and diagnostic detection. The primer comprises an upstream primer GAstV-F and a downstream primer GAstV-R, the sequences of the upstream primer GAstV-F and the downstream primer GAstV-R are respectively shown as GAstV-F / R. The kit comprises the primer. According to the GAstV-1 and GAstV-2 gene conserved regions, universal detection primers are designed, each reaction condition of an RT-PCR system is optimized, a universal one-step RT-PCR detection method for detecting GAstV is established, and the universal one-step RT-PCR detection method has the characteristics of specificity, sensitivity, rapidness, accuracy and the like, and can be used for laboratory diagnosis, customs quarantine and molecular epidemiological research of GAstV infection.

The invention relates to the field of biological products, and particularly discloses a goose astrovirus, a goose astrovirus egg yolk antibody as well as a preparation method and application of the goose astrovirus egg yolk antibody. The invention relates to a goose astrovirus, which is named as RD240629A, is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC No: 46197, and the preservation date is September 23, 2024. The goose astrovirus is named as RD240629A, and the goose astrovirus is named as RD240629A and is preserved in the China General Microbiological Culture Collection Center (CGMCC). The goose astrovirus can be used for preparing the egg yolk antibody for preventing gout of goslings, the egg yolk antibody is good in safety, no local or whole-body adverse reaction caused by antibody injection occurs, infection of the novel goose astrovirus can be effectively prevented and / or treated, and the anti-gout effect is good. The prepared egg yolk antibody is evaluated by adopting antibody titer determination and a challenge protection test, the titer of the egg yolk antibody is not lower than 1: 256, and the egg yolk antibody has a good commercial development prospect.

The invention provides a traditional Chinese medicine composition for preventing and treating gout of goslings, a traditional Chinese medicine preparation, feed, a preparation method and application, and belongs to the field of livestock breeding. The traditional Chinese medicine composition for preventing and treating gout of goslings can effectively prevent and / or treat gout of goslings caused by GAstV, and can be used for preventing and treating gout of goslings caused by goose astroviruses (the protection rate of a prevention group reaches 80%, and the protection rate of a treatment group reaches 93.33%); the growth performance of the goslings can be remarkably promoted, and the immunity of the goslings can be improved. Moreover, the traditional Chinese medicine composition for preventing and treating gout of goslings is precise and appropriate in formula, simple to prepare and remarkable in effect, and has a great popularization prospect for replacing or reducing use of antibiotics.

This invention relates to a primer set and probe set for dual real-time quantitative PCR detection of GAstV-1 and GAstV-2. The sequences of the primer set are shown in SEQ ID NO. 1-3, and the sequences of the probe set are shown in SEQ ID NO. 4-5. This invention provides a primer set, probe set, and kit for dual real-time quantitative PCR detection of GAstV-1 and GAstV-2, and establishes a detection method capable of simultaneously detecting GAstV-1 and GAstV-2. This method has advantages such as saving primer quantity, reducing detection costs, simultaneous detection, rapid detection, high efficiency, accurate quantification, high sensitivity, strong specificity, and good reproducibility. It lays the foundation for conducting molecular epidemiological surveys of GAstV-1 and GAstV-2 and subsequent scientific prevention and control of related diseases, and has significant research value.

The present invention belongs to the field of biotechnology, and discloses the use of an LMH cell line overexpressing the RPL7 gene, and a vaccine; Infecting goose astrovirus into an LMH cell line capable of overexpressing goose-derived RPL7 can, to a certain extent, promote the proliferation of goose astrovirus. As a component of the large ribosomal subunit, RPL7 directly participates in the process of mRNA to protein synthesis. GAstV mRNA interacts with RPL7 and localizes to the endoplasmic reticulum, greatly promoting the reverse transcription level of the virus, and thus promoting the replication of goose astrovirus; When it is applied to vaccine research and development, it can also obtain goose astrovirus with a high titer, which helps to reduce the cost of vaccine research and development and improve the effect of the vaccine.

The invention discloses a double-genotype goose astrovirus multi-epitope fusion protein and a preparation method and application thereof, and relates to the technical field of chemistry. Screening of GAstV positive serum; designing and synthesizing a B cell epitope of the GAstV cap protein; screening of dominant B cell epitopes of the single genotype GAstV cap protein; expression and purification of the fusion epitope all-CAP; establishing a fusion epitope ELISA antibody detection method; detecting a clinical sample; and making a detection conclusion according to a detection result. According to the double-genotype goose astrovirus multi-epitope fusion protein as well as the preparation method and the application thereof, prepared GAstV single-genotype positive goose serum is used for screening to obtain cap protein B cell epitope polypeptide with good reactivity, and the fusion epitope polypeptide all-CAP is obtained after tandem expression and purification; a universal indirect ELISA antibody detection technology which is strong in specificity, high in sensitivity and good in repeatability and can be used for common detection of GAstV-1 and GAstV-2 is established by taking the GAstV-1 and GAstV-2 as a coating antigen.

The present invention discloses a hybridoma cell line secreting monoclonal antibodies against goose astrovirus cap protein, the monoclonal antibodies, and their preparation methods and applications, belonging to the field of immunotechnology. In the present invention, mice are immunized with goose astrovirus cap protein, and specific hybridoma cell lines 7B2 and 3D4 are isolated from mouse ascites, and monoclonal antibodies are isolated and purified. Further analysis shows that the epitope information recognized by the monoclonal antibody secreted by hybridoma cell line 7B2 is 153 NTAGPESIDT 162 . Homology analysis of this epitope information is carried out among various virus strains, and it is found that this epitope information is highly conserved in goose astrovirus, indicating its potential to specifically inhibit goose astrovirus, and it can be used as a target for the development of therapeutic drugs or vaccines against goose astrovirus.

The present invention relates to a novel goose astrovirus vaccine and a preparation method and application thereof. According to the coding sequence of the CAP gene of goose astrovirus, the CAP protein fragment coding gene is determined by analyzing its conservative region and the dominant antigen epitope structure region. The CAP protein fragment is successfully expressed and purified in Escherichia coli, and a subunit vaccine of goose astrovirus is further prepared. Immunization experiments show that the prepared subunit vaccine can protect goslings, and even for goslings initially infected with the virus, it has the effect of further killing the virus in vivo. The goose astrovirus subunit vaccine of the present invention has the advantages of simple preparation method, low production cost and good immune effect, and is suitable for various poultry, such as chickens, ducks and geese, for the prevention and treatment of astrovirus.

The invention discloses a hybridoma cell strain secreting an anti-goose astrovirus cap protein monoclonal antibody, the monoclonal antibody as well as a preparation method and application of the monoclonal antibody, and belongs to the technical field of immunity. According to the invention, a goose astrovirus cap protein is utilized to immunize a mouse, specific hybridoma cell strains 7B2 and 3D4 are separated and obtained from mouse ascites, and the monoclonal antibody is separated and purified. Further analysis shows that the epitope information recognized by the monoclonal antibody secreted by the hybridoma cell strain 7B2 is 153NTAGPESIDT162, homologous analysis is carried out on the epitope information in various virus strains, it is found that the epitope information is highly conserved in the goose astrovirus, it is indicated that the epitope information has the potential of specifically inhibiting the goose astrovirus, and the epitope information can be used for preparing the goose astrovirus. The method can be used for developing goose astrovirus treatment drugs or vaccines as a target spot.

The invention discloses a hybridoma cell strain based on type 1 goose astrovirus, a construction method, a monoclonal antibody and a detection reagent. The hybridoma cell strain is a hybridoma cell strain A5A1, the preservation number of the hybridoma cell strain is CCTCC (China Center for Type Culture Collection) NO: C202505, and the hybridoma cell strain is preserved in the China Center for Type Culture Collection on March 26, 2025. According to the invention, a mouse is immunized by GAstV-1ORF2 recombinant protein expressed by a prokaryotic expression system, and a monoclonal antibody A5A1 is obtained by screening and subcloning. Indirect competitive ELISA (icELISA) for detecting the GAstV-1 antibody is developed based on the hybridoma cell strain A5A1, and the method is high in sensitivity, good in repeatability and high in specificity, only reacts with serum of GAstV-1, and does not have cross reaction with other detected pathogenic serum. A serological detection method established by the monoclonal antibody prepared by the method lays a foundation for scientific prevention and control of gout of goslings.

The invention belongs to the technical field of microorganisms, and particularly discloses a method for preparing self-assembled goose astrovirus vp27 protein-ferritin by using a baculovirus expression system, which comprises the following steps: firstly, constructing a recombinant plasmid pFastBac Dual-VP27 and a recombinant plasmid pFastBac Dual-VP27-ferritin; the preparation method comprises the following steps: respectively transforming a recombinant plasmid pFastBac Dual-VP27 and a recombinant plasmid pFastBac Dual-VP27-Ferritin into a DH10Bac competent cell, so as to obtain a recombinant positive clone bacmid-VP27 and a recombinant positive clone bacmid-VP27-Ferritin, and respectively transforming the recombinant plasmid pFastBac Dual-VP27 and the recombinant plasmid pFastBac Dual-VP27-Ferritin into a DH10Bac competent cell; the method comprises the following steps: respectively transfecting a recombinant positive clone bacmid-VP27 and a recombinant positive clone bacmid-VP27-Ferritin into an insect cell, and carrying out expression of a recombinant baculovirus, so as to obtain a recombinant baculovirus Ac-VP27 and a recombinant baculovirus Ac-VP27-Ferritin; the method comprises the following steps: respectively carrying out subculture on a recombinant baculovirus Ac-VP27 and a recombinant baculovirus Ac-VP27-Ferritin, and carrying out expression of a VP27 protein and a VP27-Ferritin protein; and then separating and purifying the VP27 protein and the VP27-Ferritin protein from the culture supernatant. According to the invention, the VP27 protein and the VP27-Ferritin protein with natural conformations can be conveniently and rapidly obtained, and the VP27 protein and the VP27-Ferritin protein act as candidate vaccines for preventing goose astrovirus infection and antigen substances for detecting antibodies of goose astrovirus.

The invention relates to the field of animal medicine detection, in particular to a goose astrovirus detection card and application thereof. The invention discloses a goose astrovirus detection card which comprises a bottom plate and immunochromatography test paper located on the bottom plate, and the immunochromatography test paper comprises a sample pad, a combination pad, a detection pad and a water absorption pad in the chromatography direction of a sample to be detected; the combination pad contains a goose astrovirus monoclonal antibody, and the goose astrovirus monoclonal antibody is labeled with a marker. The goose astrovirus detection card has the advantages of being rapid, convenient and accurate in detection and free of operation of professional experimenters.

The invention belongs to the technical field of biological detection, and relates to a detection primer group, a reagent and a kit for detecting a novel goose astrovirus (NGAstV) and a goose circovirus (GoCV) by a multi-enzyme constant-temperature rapid amplification technology (MIRA). According to the invention, two dual MIRA detection methods aiming at NGAstV and GoCV are successfully established, and in a basic dual MIRA method, target genes of the two viruses can be simultaneously and stably amplified by reacting for 25 minutes at the constant temperature of 25 DEG C; according to the fluorescent dual MIRA, the FAM and ROX labeled probes are utilized, so that the reaction time of the detection method is further shortened to 20 minutes under the condition of 39 DEG C. The two dual MIRA detection methods have good specificity, the lowest detectable concentrations of NGAstV and GoCV in dual MIRA-qPCR detection are 1 * 10 < 1 > copies / L and 1 * 10 < 2 > copies / L respectively, and the requirements of clinical rapid, convenient, efficient and accurate pathogen detection can be met.

The application discloses a different genotype goose astrovirus multiplex fluorescent PCR detection primer probe set, a kit and application, relates to the field of molecular biology; the primer probe set comprises a primer pair and a probe for detecting type 1 goose astrovirus (GoAstV-1) and type 2 goose astrovirus (GoAstV-2); the primer pair for detecting GoAstV-1 is as shown in SEQ ID NO. 1-2, and the probe is as shown in SEQ ID NO. 3; the primer pair for detecting GoAstV-2 is as shown in SEQ ID NO. 4-5, and the probe is as shown in SEQ ID NO. 6. The primer probe set disclosed by the application can realize multiplex fluorescent PCR detection of different genotypes of goose astrovirus, the detection method has good specificity, sensitivity and repeatability, and provides technical support for clinical diagnosis of GoAstV, determination of virus load and epidemiological investigation.

The invention provides a nucleic acid molecule for coding a novel goose astrovirus resisting nano antibody or application of the novel goose astrovirus resisting nano antibody coded by the nucleic acid molecule in preparation of a novel goose astrovirus resisting medicine, and belongs to the technical field of biological products. The invention discloses a nucleic acid molecule for coding a nano antibody (Nb) for resisting novel goose astrovirus (nGAstV). The nucleic acid molecule comprises a 5 '-untranslated region, a sequence for coding a signal peptide, a coding sequence of Nb, a 3'-untranslated region and a poly (A) tail which are connected in sequence. The nucleic acid molecule can efficiently and stably express Nb with immunocompetence in vivo and in vitro, experiments prove that the Nb has the activity of inhibiting replication of nGAstV, and the nGAstV is neutralized by blocking adsorption and invasion of the nGAstV to host cells. The invention provides a new way for treating the goose gout disease, and also provides a new means for developing the medicine for treating the poultry goose gout disease.

The application discloses a goose astrovirus weakly virulent vaccine candidate strain (referred to as GAstV-GXNNP120), which has a preservation number of CCTCC No:V202564 and a preservation date of September 17, 2025. Research shows that GAstV-GXNNP120 has the advantages of high virus titer, good immunoprotective effect and long antibody duration. In addition, GAstV-GXNNP120 can also be well adapted to LMH cells, and compared with culturing viruses in goose embryos, the cost of culturing viruses in the passage cells is lower, and the quality between batches is more stable. Therefore, the goose astrovirus weakly virulent vaccine candidate strain of the application can be used for further development of a goose astrovirus attenuated live vaccine, and has important significance for clinical prevention and control of goose astrovirus.

The invention aims to provide a goose astrovirus and an application of the goose astrovirus in preparation of vaccines, the goose astrovirus is named as goose astrovirus GAstV-M705, the taxonomic name of the goose astrovirus is Avastrovirus-Goose astrovirus, the goose astrovirus is preserved in the China Center for Type Culture Collection, the preservation number is CCTCC NO: V202521, and the preservation date is April 29, 2025. The strain is subjected to passage and proliferation on goose embryos and LMH cells, it is found that the strain can be stably proliferated, then the strain serves as an antigen to be prepared into the inactivated vaccine, and a challenge protection test proves that the inactivated vaccine has good immunogenicity.

The invention belongs to the technical field of veterinary biological products, and discloses an inactivated vaccine and egg yolk antibody for preventing and controlling goose astrovirus as well as a preparation method and application of the inactivated vaccine and egg yolk antibody. The invention discloses a goose astrovirus named as N-GoAstV-hs2403, the inventor obtains the goose astrovirus N-GoAstV-hs2403 with excellent immunogenicity through long-term research and separation, high-titer goose astrovirus liquid is prepared on the basis of the goose astrovirus N-GoAstV-hs2403, the titer is FAID50 = 108.50 / mL, an efficient inactivated vaccine and an egg yolk antibody are further developed, and the goose astrovirus N-GoAstV-hs2403 can be used for preparing the goose astrovirus. The titer and the production efficiency of the antibody are remarkably improved, and an innovative solution is provided for prevention and control of the goose astrovirus disease.

The invention discloses a hybridoma cell strain based on type 2 goose astrovirus, a construction method, a monoclonal antibody and a detection reagent. The hybridoma cell strain is a hybridoma cell strain A10D12, the preservation number of the hybridoma cell strain is CCTCC (China Center for Type Culture Collection) NO: C202506, and the hybridoma cell strain is preserved in the China Center for Type Culture Collection on March 26, 2025; or the hybridoma cell strain is a hybridoma cell strain A12G1, the preservation number of the hybridoma cell strain A12G1 is CCTCC NO: C202507, and the hybridoma cell strain A12G1 is preserved in the China Center for Type Culture Collection on March 26, 2025. According to the invention, purified escherichia coli expressed recombinant 2-type goose astrovirus capsid protein is used for immunizing mice, two monoclonal antibodies specifically aiming at GAstV-2ORF2 protein are obtained, and B cell epitopes recognized by the monoclonal antibodies are identified. Based on the monoclonal antibodies, an efficient detection immunological detection method for the goose astrovirus type 2 is established.

The invention discloses a VHH chain of an anti-goose astrovirus P2 protein nano antibody. The amino acid sequence of the VHH chain is as shown in SEQ ID NO: 1. Goose astrovirus is used as a research object, the goose astrovirus is inactivated to immunize alpaca, and a bacteriophage display technology is used for screening to obtain a broad-spectrum nano antibody for resisting goose astrovirus P2 protein. Through the preparation of the nano antibody for resisting the goose astrovirus P2 protein, a material is provided for establishing a broad-spectrum diagnosis method.

The invention belongs to the technical field of biology, and discloses application of an LMH cell line of an overexpressed RPL7 gene and a vaccine. Goose astrovirus is infected to an LMH cell line capable of overexpressing goose source RPL7, proliferation of the goose astrovirus can be promoted to a certain extent, RPL7 serves as a constituent part of a ribosome large subunit and directly participates in the synthesis process from mRNA to protein, GAstV mRNA and RPL7 interact and are located in an endoplasmic reticulum, the reverse transcription level of the virus is promoted to a large extent, and the effect of overexpressing the goose source RPL7 is achieved. Further, the replication of the goose astrovirus is promoted; when the goose astrovirus is applied to vaccine research and development, goose astrovirus with relatively high titer can be obtained, so that the cost of vaccine research and development can be reduced, and the effect of the vaccine can be improved.

The application discloses a type 2 GAstV capsid protein monoclonal antibody and an antigen epitope peptide thereof. The monoclonal antibody is prepared after a mouse is immunized with a type 2 goose astrovirus capsid protein GAstV Cap as an antigen and is named 1F1. The monoclonal antibody 1F1 comprises a heavy chain variable region and a light chain variable region, and the amino acid sequences are shown in SEQ ID No. 3 and SEQ ID No. 5 in sequence. In order to determine the B cell antigen epitope of the screened monoclonal antibody, a GAstV Cap truncated fusion protein is constructed, and antigen recognition epitopes are determined through indirect ELISA and Western blotting experiments, so that the monoclonal antibody is screened step by step until the shortest linear epitope recognized by the monoclonal antibody is determined, and the amino acid sequence is shown in SEQ ID No. 10. Therefore, the monoclonal antibody and the antigen epitope thereof in the application can be applied to the research on the structure and function of the GAstV capsid protein, serological diagnostic tools and the preparation of epitope vaccines.