273 results about "Foot mouth disease virus" patented technology

The present invention discloses a method for assembling foot-and-mouth disease virus empty capsids in insect cells via the alteration of acid-resistance. The method for assembling foot-and-mouth disease virus empty capsids in insect cells includes the following steps: (1) the altered P12A gene and the non-structural protein gene 3C of foot-and-mouth disease virus are introduced into bacteria via baculovirus vectors for recombination to produce recombinant rhabdovirus A; (2) the DNA of the recombinant rhabdovirus A is used to transfect the insect cells, so that the foot-and-mouth disease virus empty capsids are obtained. The method assembles the integral foot-and-mouth disease virus empty capsids in the insect cells for the first time, lays a foundation for the research and the development of gene-engineered subunit vaccines and novel diagnostic reagents.

Antiviral protease inhibitors, including peptidyl aldehydes, peptidyl α-ketoamides, peptidyl bisulfite salts, and peptidyl heterocycles, are disclosed, along with related antiviral compounds, and methods of using the same to treat or prevent viral infection and disease. The compounds possess broad-spectrum activity against viruses that belong to the picornavirus-like supercluster, which include important human and animal pathogens including noroviruses, enteroviruses, poliovirus, foot-and-mouth disease virus, hepatitis A virus, human rhinovirus (cause of common cold), human coronavirus (another cause of common cold), transmissible gastroenteritis virus, murine hepatitis virus, feline infectious peritonitis virus, and severe acute respiratory syndrome coronavirus.

This invention relates to a fusion protein used for preventing aftosa, its preparation method and application. This fusion protein contains O type foot-and-mouth disease virus main cytomembrane protein VP1 epitope, colibacillus thermolability toxin B subunit, thymus derived cell epitope and purification label.

The invention discloses a pig foot-and-mouth disease virus O-type broad spectrum multi-epitope recombination antigen and application thereof and belongs to the field of biological vaccines. The pig foot-and-mouth disease virus O-type broad spectrum multi-epitope recombination antigen adopts a strategy of an antigenized antibody, after main antigen epitopes of a plurality of strains of pig foot-and-mouth disease virus O-type are connected in series reasonably, the plurality of strains of pig foot-and-mouth disease virus O-type are coupled with a pig intravenous gamma globulin (IgG) heavy chain constant region to construct the pig foot-and-mouth disease virus O-type broad spectrum multi-epitope recombination antigen, and after ration through a Bio-Rad protein ration kit, the pig foot-and-mouth disease virus O-type broad spectrum multi-epitope recombination antigen and recombination foot-and-mouth disease virus 3D protein are matched to prepare the vaccines. Animal immunity testing results show that the vaccines can stimulate an organism to generate high-titer protective antibodies when the vaccines are used independently or matched with the recombination foot-and-mouth disease virus 3D protein to be used, an antibody level is higher than a national standard, and good application prospects are achieved.

This application is directed generally to foot-and-mouth disease virus (FMDV) 3C proteases that have been modified by mutating a polynucleotide sequence coding for the FMDV 3C protease. The modified FMDV proteases exhibit proteolytic activity on FMDV P1 precursor protein and exhibit a reduction in one or more toxic or inhibitory properties associated with an unmodified FMDV 3C protease on a host cell used to recombinantly produce it. Vectors carrying polynucleotides encoding modified FMDV 3C protease sequences can induce production of FMDV virus-like particles in a host cell when expressed in the host cell. The modified FMDV 3C proteases can generally be used to produce immunogenic FMDV preparations capable of inducing an immune response against FMDV.

The invention relates to a multiplex PCR (polymerase chain reaction) primer, a probe and a gene chip for detecting the bluetongue virus, foot and mouth disease virus and bovine viral diarrhea virus. The multiplex PCR primer and probe have the nucleotide sequences shown by SEQ ID No.1 to SEQ ID and No.9. The gene chip comprises a solid-phase carrier, a sample application quality control probe, a positive hybrid quality control probe and a multiplex PCR primer for detecting the bluetongue virus, foot and mouth disease virus and bovine viral diarrhea virus and the corresponding probe. In the invention, the forward primers of three viruses are marked with fluorescence, a gene chip detection technology carrying three viruses in animal fur is established based on multiplex RT-PCR (reverse transcription-polymerase chain reaction), and the RNA virus in the fur can be sensitively and specifically detected with high flux; the three viruses are screened at the same time in detection once, and the situation that a specific method is required for each virus before is changed, thereby saving the diagnosis time, meeting the needs for quick detection of mass imported/exported fur samples of the exit-entry inspection and quarantine departments and the fur import and export enterprises, and realizing relatively high application values.

The invention discloses an O type foot-and-mouth disease virus variant as well as a coding gene and application thereof. In the invention, an O type foot-and-mouth disease virus pan-Asia strain O/YS/CHA/05 is firstly separated out, the nucleotide sequence of the O type foot-and-mouth disease virus pan-Asia strain O/YS/CHA/05 SEQ ID NO: 1, and the amino acid sequence is SEQ ID NO: 2. In comparison with a VP1 amino acid sequence, the virus strain has 7 variable sites, five of which are centralized in a G-H ring. Mutation of the sites ensures that the virus variant has the capability of escaping from host immunity so as to have the superiority for becoming a popular virus strain. Therefore, the variant can be employed to prepare an inactivated vaccine for prevention and treatment of the variant and relevant strains, dominant antigen epitope of the variant can be employed to prepare a synthetic peptide vaccine, and the variant can be employed to develop novel O type foot-and-mouth disease virus vaccines such as VLP vaccine and the like. Therefore, the invention has important value in controlling the popularity of O/YS/CHA/05 and relevant variable strains.

The invention belongs to the technical field of biological detection, and discloses HRM detecting primers and a method for distinguishing the foot-mouth disease virus (FMDV) and the Seneca Valley virus (SVV). The primers have the sequences shown in SEQ ID NO:1 and SEQ ID NO:2 and are high in specificity. By means of the primers, PCR amplification is conducted on the FMDV and the SVV, then fluorescent data is collected by monitoring the combination situation of double-chain DNA fluorescent dyes and PCR amplification products in the temperature rise process in real time, and the FMDV and the SVV are distinguished according to the difference of two dissolution curves; the two gene types can be distinguished after PCR amplification is conducted through the primers, it takes people only 3 hours for the whole operation process, no virus cell culture is needed, and the type distinguishing time is greatly shortened; expanses are low, no specific probe is needed, and fluorescent saturated dyes are low in price and easy to obtain; accuracy, specificity and repeatability are high, analysis can be accurately and rapidly conducted at high throughput, and the primers and method are easy to apply and popularize in clinical practices.

The invention discloses a foot-and-mouth disease virus (FMDV) resistant monoclonal antibody and an identified epitope and application thereof, and belongs to the field of prevention and control of the FMDV. The microbial collection number of a hybridoma cell line, which can secrete the neutralizing monoclonal antibody resisting to Asia-1 FMDV, is CGMCC No.2692; and the microbial collection number of the hybridoma cell line, which can secrete the neutralizing monoclonal antibody resisting to O-type FMDV, is CGMCC No.2691. The invention also discloses amino acid sequences of a conformational neutralizing epitope of the Asia-1 FMDV VP1 protein and a linear neutralizing epitope of the O-type FMDV VP1 protein which are identified by the two monoclonal antibodies respectively. In-vitro neutralization tests and in-vivo animal protection tests show that both the two monoclonal antibodies have excellent passive immunity effect, can be applied to emergency prevention of the FMDV and have excellent immunity effect on the passive immunity of the FMDV.

The invention discloses a foot and mouth disease virus recombinant virus sample particle as well as a preparation method and application thereof. The recombinant virus sample particle is commonly assembled and expressed by components in a DNA molecule composition. The DNA molecule composition contains an O-type foot and mouth disease VP0 gene, an O-type foot and mouth disease VP1 gene and an O-type foot and mouth disease VP3 gene and further contains a green fluorescent protein gene. By virtue of the property that FMDV VLPs is self-assembled through VP0, VP1 and VP3, a construction method of a baculovirus recombinant vector is improved, a green fluorescent protein label is added into the vector, and FMDV VLPs is successfully prepared by virtue of a pFBDM Bac-to-Bac system, so that a theoretical foundation is laid for the further development of safe and efficient O-type FMDV genetic engineering vaccines.

The invention provides a type O foot-and-mouth disease virus mutant and a preparation method and application thereof, and belongs to the technical field of vaccine candidate strains. According to thetype O foot-and-mouth disease virus mutant disclosed by the invention, an rHN virus strain is used as a female parent virus strain, and the following amino acids in a G-H ring of VP3 protein are mutated: the 173rd aspartic acid is mutated into asparagine, the 174th valine is mutated into glutamic acid and the 179th asparagine is mutated into cysteine. The virus mutant has heredity stability and obtains the capacity of caveolin for performing mediated infestation on CHO-K1cells. The result of detecting the cross neutralization capacity of immune positive serum indicates that compared with a female parent virus strain, the virus mutant has the advantages that the cross protection capacity of the virus mutant for inducing organisms to produce foot-and-mouth disease virus neutralizing antibodies is notably improved, and the virus mutant shows excellent antigen broad spectrum properties. Vaccines prepared through inactivation of the virus mutant can be used for preventing infection with type O foot-and-mouth disease viruses.

Based on a highly conserved domain of a foot-and-mouth disease virus 3D protein coding gene, a vesicular stomatitis virus N protein coding gene and a swine vesicular disease virus VP1 protein coding gene, the invention designs a specific primer and a probe and develops a multiple fluorescence RT-PCR detection method used for simultaneously detecting the three animal viruses. The detection method can detect 102 copied plasmids containing target amplification sequences in 1 h. The method has high sensitivity and good specificity, and can achieve rapid high-throughput fluorescence RT-PCR detection for single-virus infection or mixed infection by a plurality of viruses.

Disclosed are a method for preparing and virus type definition test paper for foot and mouth disease and the using method. The test paper comprises O, A, Asia l, C four kinds of serum detecting test paper. The test paper comprises a PVC substrate plate, nitrocellulose membrane, an absorbing pad, a golden mark pad and a sample pad. The PVC substrate plate is installed at the most bottom layer; the nitrocellulose membrane is set on the middle part of the substrate plate; the absorbing pad is stuck on the left end of the nitrocellulose membrane, and the golden mark pad is set on the right end of the nitrocellulose membrane; the sample pad is set on the right upper end of the golden pad. The invention can identify the foot and mouth virus O, A, Asia l and C four serum type.

The invention relates to an O-type foot-and-mouth disease virus antibody chemiluminescence detection kit. The kit includes an polystyrene board coated by antigen, a monoclonal antibody, a standard substance, a luminescence substrate solution A and a luminescence substrate solution B. The polystyrene board coated by antigen is an opaque polystyrene plate coated by an O-type foot-and-mouth disease virus RE2 recombinant protein; the monoclonal antibody is a horseradish peroxidase labeled O-type foot-and-mouth virus monoclonal antibody; the luminescence substrate solution A comprises a Lumino, hydroxycoumarin, gallic acid, a Tris-Hcl buffer and water; the standard substance is foot-and-mouth virus O-type antibody respectively diluted buy a calibration diluent, wherein the dilution is 0NU / mL, 2NU / mL, 5NU / mL, 10NU / mL, 30NU / mL and 60NU/ mL respectively. The kit of the invention can be used for detecting O-type foot-and-mouth disease virus antibody, has the advantages of high sensitivity and wide detection range, and realizes the quantitative detection of foot-and-mouth disease antibody.

The invention discloses a primer combination for identifying foot-and-mouth disease virus and vesicular stomatitis virus and application thereof. The primer combination is composed of a primer set I and a primer set II. The primer set I is composed of primers FMDV-F3, FMDV-B3, FMDV-FIP and FMDV-BIP which are sequentially disclosed as Sequence 1-4. The primer set II is composed of primers VSV-F3, VSV-B3, VSV-FIP and VSV-BIP which are sequentially disclosed as Sequence 5-8. The invention also discloses application of the primer combination in identifying foot-and-mouth disease virus and vesicular stomatitis virus, application in identifying whether a virus to be detected is foot-and-mouth disease virus or vesicular stomatitis virus, and application in identifying whether a sample to be detected is infected by foot-and-mouth disease virus and/or vesicular stomatitis virus. The duplex RT-LAMP (reverse transcription-loop-mediated isothermal amplification) method established by the invention is a simple quick low-cost diagnosis method, can be used for grass-root and field quarantine inspection under poor conditions, and is also suitable for large-scale epidemiological survey.

The invention belongs to the technical field of bovine virus detection and particularly relates to a foot and mouth disease virus and vesicular stomatitis virus identifying duplex fluorescence RT-LAMP (loop-mediated isothermal amplification) detection primer group, a kit and application thereof. The foot and mouth disease virus and vesicular stomatitis virus identifying duplex fluorescence RT-LAMP detection primer group comprises two groups of specific primers, wherein one group is FMDV-F3, FMDV-B3, FMDV-FIP (F1c-F2) and FMDV-BIP (B1c-B2), and the other group is VSV-F3, VSV-B3, VSV-FIP (F1c-F2) and VSV-BIP (B1c-B2), and sequences of the two groups are shown as SEQ ID NO.1 and SEQ ID NO.8 respectively. An established foot and mouth disease virus and vesicular stomatitis virus identifying duplex fluorescence RT-LAMP method has advantages of simplicity, convenience, quickness, specificity, sensitivity and the like and can be used for FMDV (foot and mouth disease virus) and VSV (vesicular stomatitis virus) clinical detection and epidemiological investigation. The FMDV and VSV duplex fluorescence RT-LAMP method is a simple, quick and low-cost diagnosis method and suitable for large-scale epidemiological investigation.

The invention provides a foot and mouth disease virus antigen polypeptide, a foot and mouth disease virus fusion antigen polypeptide and a foot and mouth disease virus vaccine containing the antigen polypeptide and/or the fusion antigen polypeptide. The invention further provides application of the antigen polypeptide, the fusion antigen polypeptide and the vaccine in prevention and control of foot and mouth disease virus infection. The foot and mouth disease virus antigen polypeptide, the fusion antigen polypeptide and the vaccine have broad-spectrum immunogenicity, and can generate good immunogenicity on different foot and mouth disease viruses and variant strains thereof.

The invention discloses a gene chip and a detection method for detecting FMDV, VSV, SVDV, PPRV and BTV. The detection method comprises the step of detecting foot and mouth disease viruses (type A, Asian type I and type O), vesicular stomatitis virus, swine vesicular disease virus, Peste des petits ruminants virus and bluetongue virus. The method comprises the following specific steps: designing a PCR primer by virtue of sequence analysis of standard strain genome, and performing cloning and sequence analysis on target genes; designing a specific probe, and simultaneously detecting the foot and mouth disease viruses, vesicular stomatitis virus, swine vesicular disease virus, Peste des petits ruminants virus and bluetongue virus. The invention aims at establishing a method for detecting the foot and mouth disease viruses, vesicular stomatitis virus, swine vesicular disease virus, Peste des petits ruminants virus and bluetongue virus by adopting a microarray chip which is high in sensitivity and high in specificity and through which the time and labor are saved and the result is easily observed.

The invention relates to a recombinant vaccine strain for foot-and-mouth disease type A, which is constructed by a gene recombination technology and featured with high valence, high antigen compatibility and high immune protective rate, as well as a preparation method thereof and an application thereof. An antigen nucleotide sequence of the vaccine strain is shown by SEQ ID NO. 1; a rescuing system is an efficient eukaryotic plasmid, which is manually constructed and can express an accurate foot-and-mouth disease virus genome RNA; therefore, a foot-and-mouth disease recombinant virus can be constructed and prepared; through the adoption of the plasmid, the vaccine strain with high titer and good antigen compatibility can be prepared so as to prepare inactivated vaccines; after pigs and cattle are immunized, bodies can be effectively stimulated to generate immune responses and immune protective effects on the pig and cattle bodies can be provided; an A type AISA lineage of strain is challenged with 10000 times of 50% cattle infective dose (BID50), so that the immune protective rate reaches 100% and 50% protective dose (PD50) is 10.81-13.59; and the vaccine can be used for preventing and controlling foot-and-mouth diseases type A in China and neighboring countries.

The invention discloses a cattle food-and-mouth disease virus A type synthetic peptide. The cattle food-and-mouth disease virus A type synthetic peptide has the following amino acid sequence: acetyl-YDLDF EALKP HFKSL GQTIT PADKS PPS VYNGT CKYSA PATRR GDLGS LAARL AACLP ASFNY GAIRA T-amide. The cattle food-and-mouth disease virus A type synthetic peptide can be used as an effective novel vaccine for the A type food-and-mouth disease in production practice; simultaneously, the realization of the test provides a certain foundation for further perfecting the construction of a food-and-mouth disease virus A type synthetic peptide vaccine and the construction of other food-and-mouth disease subtype synthetic peptide vaccines; and the new ideal proposed in the research of the synthetic peptide vaccine and the construction of a new method of the synthetic peptide vaccine provide theoretical foundation and technical support for further perfecting the development and research of the multiple antigenic peptide, multivalent peptides and joint peptide vaccines in future.

A kind of siRNAs able to suppress the copy and infection of foot-and-mouth disease virus in cell and animal individual and its preparing process and application are disclosed. The 1D protein gene and 3D polymerase gene are chosen as the interference targets from the genome of foot-and-mouth disease virus. The human recombinant adenovirus Ad5 is designed and used as the carrier of three siRNAs. The target of 21VPI siRNA and 63VPI siRNA is said 1D gene. The target of 56POL siRNA is 3D gene.

The invention provides a recombinant phage double expression vector and application, relating to the technical field of biology. The recombinant phage double expression vector is a recombinant double expression vector obtained after inserting a target gene 1 into the downstream part of a gene p10B in a T7 phage genome and inserting a eukaryotic expression cassette containing a target gene 2 into a non-coding region in the T7 phage genome. The recombinant phage double expression vector has high stability, can display the protein coded by the target gene 1 on the surface of the phage, simultaneously can transfer the eukaryotic expression cassette into an immune cell to achieve eukaryotic expression of the target gene 2, and can serve as a common platform for expression of various epitopes, namely polypeptides. Immune efficacy detection finds that bodies can generate high-level VP1 structural protein antibodies against foot and mouth disease viruses after being immunized by the vaccine, and the antibodies have long duration.

The invention discloses preparation and application of a monoclonal antibody of an A-type foot and mouth disease virus (FMDV) and belongs to the field of prevention and treatment of foot and mouth diseases. According to the preparation and the application, a hybridoma cell line secreting the neutralizing monoclonal antibody of the A-type FMDV is constructed, the microbiological collection number of the hybridoma cell line is CGMCC 12049, and the secreted monoclonal antibody is an A-type FMDV serotype specific monoclonal antibody and has high-strength neutralizing ability; and a detection method for the A-type FMDV antibody is created by using the monoclonal antibody. The monoclonal antibody prepared by the invention shows good competitive ability with strongly-positive, weakly-positive and suspected serum of the A-type FMDV in the detection method for the A-type FMDV antibody and is free of competitiveness with strongly-positive serum of an O-type FMDV, strongly-positive serum of an Asia1-type FMDV and negative serum of an FMDV, so that the monoclonal antibody can be used for specifically discriminating the negative and positive serum of the A-type FMDV and has an important application prospect in aspects such as the diagnosis, prevention and treatment of the A-type foot and mouth diseases.