31 results about "Fmd virus" patented technology

The present invention relates to modified poxviral vectors and to methods of making and using the same. In particular, the invention relates to recombinant avipox that expresses gene products of foot and mouth disease virus (FMDV), and to compositions or vaccines that elicit immune responses directed to FMDV gene products and which can confer protective immunity against infection by FMDV.

The invention provides a chimeric protein, a virus-like particle and application thereof and belongs to the field of molecular biology. The chimeric protein is prepared by the following steps: replacing one, two or three of four sites in a porcine circovirus 2 type Cap protein by a main B cell epitope of a porcine O-shaped foot-and-mouth disease virus VP1 protein to obtain 58th-66th amino acid residues, 72nd-94th amino acid residues, 122nd-147th amino acid residues and 162nd-197th amino acid residues. The chimeric protein can form the chimeric virus-like particle after soluble expression, shows the main B cell epitope of the porcine O-shaped foot-and-mouth disease virus VP1 protein on the surface of the chimeric virus-like particle, has very good immunogenicity to both PCV2 and FMDV and can generate a high antibody to the PCV2 and the FMDV, by once immune.

The invention provides an antigen spectrum expanded O-type foot and mouth disease virus strain as well as a construction method and application thereof, and belongs to the technical field of veterinary drug biological products. The rHN / NXVP1 / G-H takes a recombinant virus rHN / NXVP1 as a skeleton and is embedded with a G-H ring antigen epitope gene of O / HB / HK / 99; the recombinant virus is obtained by embedding a VP1 gene of O / NXYCh / CHA / 2018 on the basis of O / HN / CHA / 93. The O / HN / CHA / 93 and the rHN / NXVP1 are not matched with Cathay pedigree virus antigens, the rHN / NXVP1 / G-H can be highly matched with PanAsia, Mya98 and Ind-2001 pedigree virus antigens and can also be matched with the Cathay pedigree virus antigens, the antigen spectrum of the foot and mouth disease virus is expanded through replacement of a G-H ring, and the prepared inactivated vaccine can be used for preventing and controlling O-type multi-pedigree FMDV prevalence.

The present invention belongs to the field of biotechnology, and is especially one kind of recombinant live carrier vaccine co-expressing pig gyrate virus and foot and mouth disease virus. The present invention aims at providing two recombinant chicken pox viruses to co-express capsid protein ORF2 or its fusion protein ORF2-ORF1 of the China epidemic strain of type-II pig gyrate virus and the structural protein precursor P1 of the China epidemic strain NY00 of type-O foot and mouth disease virus, and for serving as the duplex live carrier gene engineering vaccine to prevent PCV2 and FMDV infection. The two recombinant chicken pox viruses can stimulate mouse to generate effective body fluid immunity and cell immunity, and are safe to experimental animals without any pathological phenomenon.

The invention relates to the technical field of molecular biology, in particular to a whole genome sequence of a foot and mouth disease virus artificial attenuated virus strain O / Akesu / 58 CE39B, and aprimer and application thereof. The whole genome sequence of the foot and mouth disease virus artificial attenuated virus strain O / Akesu / 58 CE39B is shown in SEQ ID No.1. The whole genome sequence ofthe foot and mouth disease virus artificial attenuated virus strain O / Akesu / 58 CE39B is disclosed for the first time, facilitates virus reverse genetic research, lays the foundation for further research of the proliferative mechanism, the virus titer, the virulence mechanism and FMDV marker vaccines of FMDV in cells, and facilitates researching of virus evolution and virulence, molecular epidemiology and the like. The foot and mouth disease virus artificial attenuated virus strain O / Akesu / 58 CE39B has good safety and effectiveness in virus rescue, and is expected to serve as a genetic engineering vaccine of the next generation to be used for foot-and-mouth disease prevention and control, and the whole genome sequence has the wide research and industrialization prospects.

The present invention provides a yellow fever virus vector, in which an exogenous polypeptide expression module is inserted into its genome, and the expression module has in sequence from 5' to 3': (a) a release element at the 5' end; (b) an exogenous polypeptide encoding The gene element of source polypeptide; And (c) 3 ' terminal release element, described release element is selected from: the nucleotide sequence of coding foot-and-mouth disease virus autohydrolase, the nucleotide sequence of coding signal peptide hydrolase substrate, and its combination. The carrier can provide antigenic polypeptides of viruses and tumors in host cells, thereby triggering immune responses against viruses and tumors.

The invention provides a foot-and-mouth disease O-type PanAsia-2 pedigree reserve vaccine strain as well as a construction method and application thereof, and belongs to the technical field of veterinary drug biological products. The foot-and-mouth disease recombinant virus rHN / TUR09 / VP1 is obtained by taking an O / HN / CHA / 93 virus strain as a skeleton and embedding a VP1 gene of O / TURR / 5 / 2009. The rHN / TUR09 / VP1 not only is highly matched with antigens of a foot-and-mouth disease O-type PanAsia pedigree strain and an Ind-2001 pedigree strain which are relatively close to the genetic relationship, but also is matched with antigens of a foot-and-mouth disease O-type Cathay pedigree strain and a Mya-98 pedigree strain which are relatively far from the genetic relationship, so that the rHN / TUR09 / VP1 can be used for well immunizing, preventing and controlling the prevalence of the current O-type multi-pedigree FMD virus strain in China; and the strain can be used as a strategic reserve vaccine strain for effective prevention and control of O / Panaa-2 pedigree foot-and-mouth disease in border areas of China.

Compositions for prevention of Foot and Mouth Disease (FMD) are provided, comprising an antigen component in the amount equivalent to 0.5-20 μg FMD virus and an adjuvant component comprising oil, an immunostimulatory oligonucleotide, and a polycationic carrier. Methods of using the composition, as well as the methods of reducing FMD persistence are also provided.

An antiviral antisense composition and method for treating foot-and-mouth disease virus (FMDV) in veterinary animals is disclosed. The composition contains an antisense compound that has a sequence effective to target at least 12 contiguous bases of an FMDV RNA sequence within a region of the positive-strand genomic RNA defined by SEQ ID NO: 25, and preferably, one of the viral sequences within SEQ ID NO:25 identified by SEQ ID NOS: 26-28. The composition is administered in a therapeutically effective amount in treating FMDV.

The invention discloses a triple inactivated vaccine for pigs and a preparation method and application of the triple inactivated vaccine. The triple inactivated vaccine for pigs comprises recombinantbaculovirus for expressing an O-type FMDV VP1 gene, swine senecavirus, H3N2 subtype swine influenza virus and an immunologic adjuvant. The invention also discloses a method for preparing the triple inactivated vaccine for pigs. The method comprises the following steps: (1) amplifying and inactivating the recombinant baculovirus for expressing the O-type FMDV VP1 gene; (2) mixing the inactivated recombinant baculovirus fluid for expressing the O-type FMDV VP1 gene, the inactivated swine senecavirus fluid and the H3N2 subtype swine influenza virus to obtain a water phase; (3) heating the immunologic adjuvant to obtain an oil phase; and (4) adding the oil phase into the water phase, mixing and emulsifying. Immune protective efficacy detection results prove that the prepared triple inactivatedvaccine for pigs can effectively prevent and treat O-type foot-and-mouth disease virus, swine senecavirus and H3N2 subtype swine influenza virus simultaneously.

The invention provides monoclonal antibodies 6G6 and 7D7 which can specifically bind with different genetic topology O-type foot-and-mouth disease (FMD) viruses, and an ELISA detection kit prepared from the monoclonal antibodies 6G6 and 7D7 can be used for carrying out high-sensitivity detection on the different genetic topology O-type FMD viruses.

The invention relates to the technical field of viruses in biotechnology, in particular to a mutant foot-and-mouth disease virus infectious clone, and also relates to preparation and application of the infectious clone. Lysine at the 51st, 54th, 110th and 159th sites of 3C protein is mutated into arginine through a site-specific mutagenesis technology. According to the constructed mutant infectious clone FMDV IC-K5154110159R, SUMOylation modification sites of the 3C protein of the infectious clone are mutated, the infectious clone can be proliferated, and a powerful tool is provided for detecting and researching FMDV infection and SUMOylation modification at the virus level.

A multiplex-PCR (polymerase chain reaction) kit for detecting six viruses of sheep and goats simultaneously comprises six pairs of specific primers, wherein the primers used for detection of the bluetongue virus are BTV-F and BTV-R respectively; the primers for detection of the foot and mouth disease virus are FMDV-F and FMDV-R respectively; the primers for detection of the peste des petits ruminants virus are PPRV-F and PPRV-R respectively; the primers for detection of the sheep pox virus are SPPV-F and SPPV-R respectively; the primers for detection of the goat pox virus are GTPV-F and GTPV-R respectively; the primers for detection of the sore mouth disease virus are ORFV-F and ORFV-R respectively. The sequences of the primers are nucleotide sequences shown in SEQ ID NO: 1-12. The multiplex-PCR kit can be used for detecting nucleic acid of the six viruses including the bluetongue virus, the foot and mouth disease virus, the peste des petits ruminants virus, the sheep pox virus, the goat pox virus and the sore mouth disease virus in sheep and goat clinical samples and has the characteristics of high sensitivity, high specificity, simple operation and reduction of detection time.

According to simulation results of a computer software, SAT2 FMDV B cell epitope (VYTKAAAAIRGDRAALAAKYADTNHTLPPTFNFGYVTVDK) is embedded in a Loop2 region of porcine parvovirus PPV VP2 protein, and a Tcell epitope (NVQEGRRKHTDVAFLLDRST) is embedded in an N end. A chimeric gene GS1923OP is synthesized in an optimized mode according to insect cell tropism and cloned onto a vector pFastBacTMDual, a competent cell DH10Bac is transformed to obtain recombinant bacmid rBacmidNT1L2B to transfect a sf9 cell, recombinant baculovirus rBacNT1L2B is harvested and used to infect a HF cell, an indirect immunofluorescence IFA needle is used to detect a chimeric epitope and VP2 protein, and specific fluorescence is achieved. A We stern-blot needle is used to detect the B cell epitope, the T cell epitope and the VP2 protein, and a specific band appears at 67KDa. Transmission electron microscope (TEM) observation shows that the recombinant protein NT1L2B can self-assemble into virus-like particles. The preparation and application can be applied to prepare a reserve vaccine for South Africa foot-and-mouth disease type 2, and can achieve the purpose of simultaneously preventing South Africa foot-and-mouth disease type 2 and Porcine parvovirus infection on the basis.

The invention discloses a foot-and-mouth disease virus-like particle, a preparation method, a vaccine composition and application. The vaccine composition of the present invention is composed of O-type foot-and-mouth disease virus-like particles composed of foot-and-mouth disease virus structural proteins VP4, VP2, VP3, and VP1 and / or Asian type 1 foot-and-mouth disease virus-like particles and / or foot-and-mouth disease virus-like particles composed of foot-and-mouth disease structural proteins VPO, VP3, and VP1 The type A foot-and-mouth disease virus-like particles composed of structural proteins VP0, VP3, and VP1 are composed of an adjuvant, which has a stable structure and reasonable components. It can quickly form specific antibodies, significantly increase the duration of immunity, and maintain long-term immune protection.

The present application relates to a method for preparing a foot-and-mouth disease (FMD) vaccine, comprising the following steps: (i) obtaining cell culture media containing FMD virus; (ii) separating and purifying the cell culture media containing FMD virus by an integrated filtration system with two membranes in combination; and (iii) collecting the concentrated solution containing FMD virus obtained in step (ii). The present application also relates to an FMD vaccine prepared by the method described herein and use thereof in the manufacture of a medicament for preventing animal FMD. The present application further relates to an apparatus for preparing an FMD vaccine, which comprises an integrated filtration system with two membranes in combination.

The invention provides a Seneca recombinant virus of recombinant O-type foot-and-mouth disease virus epitope gene, a recombinant vaccine, a preparation method and application thereof, and relates to the technical field of genetic engineering. The present invention obtains the full-length cDNA of SVV / FJ / 001 strain, and performs deletion and mutation transformation on its 5'UTR, and at the same time fuses the recombinant epitope gene of O-type FMDV into the SVA cDNA to construct the recombinant foot-and-mouth disease antigen epitope The Seneca recombinant virus, which can express the fused foot-and-mouth disease B-cell epitope and T-cell epitope, and the expression product has good reactogenicity, and the pathogenicity of the recombinant virus is significantly reduced or even non-pathogenic to pigs It can significantly improve the biological safety of the strain; the prepared inactivated vaccine has good immunogenicity, and can effectively stimulate SVA neutralizing antibodies while also producing specific antibodies against FMDV, which can be used for Seneca virus and Control of one or more non-Seneca viruses.

The invention relates to the application of buquinal in the preparation of medicines for preventing foot-and-mouth disease virus infection, and belongs to the technical field of veterinary medicines. The application of the invention can provide a class of highly efficient, safe and quality-controllable anti-foot-and-mouth disease virus medicine for further controlling the spread of foot-and-mouth disease.

The invention discloses a foot-and-mouth disease virus non-structural protein monoclonal antibody 7D7, a foot-and-mouth disease virus non-structural protein antibody detection kit containing the monoclonal antibody and an application thereof. The antibody detection kit provided by the invention has high sensitivity and can obviously widen the linear range of detection; the detection is not limited by animal species, and is suitable for large-scale screening of foot-and-mouth disease diseases of artiodactyl animals. The vaccine prepared by the conjugate containing the monoclonal antibody 7D7 can remove non-structural proteins, and can avoid the interference of FMDV non-structural protein antibodies after immunizing animals, thereby accurately distinguishing infected animals from immunized animals.

The invention relates to a double-link rapid detection card for detecting and distinguishing O-type and A-type foot-and-mouth disease viruses and a preparation method thereof. The detection card uses the principle of double-antibody sandwich ELISA to detect antigens. The monoclonal antibody mAbI that can specifically recognize the O-type foot-and-mouth disease virus and the monoclonal antibody mAbII that can specifically recognize the A-type foot-and-mouth disease virus are sprayed onto the detection membrane (nitrocellulose membrane) ) at the T1 and T2 detection line positions to prepare a detection imprint; spray goat or rabbit anti-mouse IgG or SPA to the C line position of the quality control area of ​​the detection membrane to prepare a quality control imprint. The gold-labeled antibody fiber layer is divided into upper and lower layers, which are respectively adsorbed with colloidal gold-labeled monoclonal antibody mAbIII that can specifically recognize type O foot-and-mouth disease virus and monoclonal antibody mAbIV that can specifically recognize type A foot-and-mouth disease virus. The test strip can be used for rapid detection of O-type and A-type foot-and-mouth disease virus (FMDV) infection. By applying the invention, the operation is simple and fast, the result is clear and easy to argue, and is suitable for popularization at the grassroots level.

The invention discloses a monoclonal antibody blocking enzyme-linked immunosorbent assay (ELISA) kit and a method for detecting the nonstructural protein (NSP) antibody of a foot-and-mouth disease virus (FMDV) (FMD NSP B-ELISA); the kit comprises ELISA reaction plates, serum diluent, 25 times concentrated detergent, substrate solution, 100* concentrated ELISA detecting antibody, stop buffer, positive control serum and negative control serum; the ELISA reaction plates are two 96-pore high-affinity ELISA reaction plates, firstly 6* groups of amino acid monoclonal antibody or NSP 2C polyclonal antibody, and then FMDV 3ABC or 2C3AB NSP which is expressed by pronucleus and is provided with 6* groups of amino acid labels is captured through the monoclonal antibody or the polyclonal antibody; and compared with other similar kits, the method has higher coincidence rate and higher positive serum detection rate, and is applicable to detecting the serum of cattle, sheep, pigs and other susceptible animals.

The invention discloses a recombinant foot-and-mouth disease virus non-toxic strain with heat-resistant phenotypic stable inheritance and a negative marker and an O / A type foot-and-mouth disease bivalent inactivated vaccine. According to the invention, FMDV virus cDNA infectious clone plasmids are constructed, and the plasmids carry a molecular determinant of virus capsid heat-resistant phenotypes, a molecular factor losing replication capacity in vivo, 3A and 3B protein epitope missing negative marker factors and Pst I restriction enzyme cutting sites introduced to two sides of a P1 coding region. The capsid protein coding region of any epidemic strain can be replaced to quickly construct and save the heat-stable and labeled foot-and-mouth disease virus non-toxic strain, and the non-toxicstrain can be used as a foot-and-mouth disease inactivated vaccine seed virus. The universal plasmids are used for constructing two recombinant viruses aiming at two current dominant epidemic strainsand preparing the O / A type foot-and-mouth disease bivalent inactivated vaccine by using the recombinant viruses, and the vaccine is used for immunizing animals to induce high-level neutralizing antibodies and generate immune protection; and the vaccine is used for inoculating animals for antibody detection, and differential diagnosis of the vaccination animals and naturally-infected animals can be realized.

Early protection of susceptible animals against foot and mouth disease (FMD) may be achieved by inoculating the animals with a vaccine comprising an interferon DNA sequence. One day after inoculation, animals have been found protected from challenge with virulent foot and mouth disease virus. Co-administration with an effective foot and mouth disease virus vaccine provides protection prior to the development of specific immunity, a feature especially desireable during a FMD outbreak.

The present invention relates to the technical field of molecular biology, and relates to an artificial attenuated strain O / Akesu / 58 CE39B genome sequence, primers and applications thereof. The artificially attenuated strain O / Akesu / 58 CE39B genome sequence is as follows: Shown in SEQ ID No.1. The present invention discloses the full genome sequence of the artificially attenuated strain O / Akesu / 58 CE39B of foot-and-mouth disease virus for the first time, which is helpful for the reverse genetics research of the virus, and is for further research on the proliferation mechanism, virus titer and pathogenic mechanism of FMDV in cells It lays the foundation for FMDV-marked vaccines, which is conducive to the study of virus evolution, pathogenicity, and molecular epidemiology; and the artificially attenuated strain of foot-and-mouth disease virus O / Akesu / 58 CE39B rescued virus has good safety and effectiveness, and is expected to be used as a vaccine. The next-generation genetically engineered vaccine is used for the prevention and control of foot-and-mouth disease, and has broad prospects for research and industrialization.

The invention discloses a method for acquiring anti-FMD (foot-and-mouth disease) transgenic goats or pigs by knocking out FMD virus receptor integrin beta6 subunit genes, which comprises the following steps: transfecting embryo fibroblasts of goats or pigs with linearized targeting carriers having report-gene-mutated integrin beta6 subunit genes; carrying out homologous recombination to obtain nuclear donor cells with the integrin beta6 subunit genes replaced; introducing nuclear donor nuclei into enucleated oocytes of goats or pigs to acquire reconstructed embryos; implanting the reconstructed embryos into wombs of surrogate goats or pigs to acquire a filial generation which belongs to heterozygote transgenic goats or pigs with the integrin beta6 subunit genes knocked out; and carrying out secondary targeting and somatocyte cloning on the heterozygote transgenic goats or pigs to acquire homozygote transgenic goats or pigs. The transgenic goats or pigs acquired by the method carry out low-expression or non-expression on the FMD virus receptor integrin beta6 subunits, obviously reduce the FMD virus infection rate and have the anti-FMD capability.

The invention relates to a duck tembusu virus transient transfection replicon carrying ranilla luciferase as well as a construction method and application of the duck tembusu virus transient transfection replicon. The replicon is characterized in that a ranilla luciferase reporter gene is linked after a tembusu virus deletes an encoding structure gene and the sequences of 38 amino acids in front of N-terminal of C protein are encoded; a sequence FMDV2A for encoding self-splicing peptide of a foot and mouth disease virus is inserted behind Rluc; an eukaryotic promoter CMV and a prokaryote promoter T7 are added to the upstream of the 5' UTR sequence of the tembusu virus; hepatitis delta virus ribozyme sequence HDVr and SV40poly(A) sequence are added to the downstream of 3' UTR; the obtainedreplicon can accurately and quantitatively reflect replication conditions of the replicon in cells and can be conveniently applied to research of replication mechanisms of the tembusu virus or even other flaviviruses, antiviral drug screening, virus and protein interaction and the like.