33 results about "Senecavirus" patented technology

The invention belongs to the technical field of biological medicine, and particularly relates to application of anethomycin in preparation of medicine for inhibiting cysteine protease activity. The invention discovers that anethomycin has the function of inhibiting the activity of virus cysteine protease, especially 3C or 3CL protease, for the first time. Molecular docking proves that anethomycin can effectively bind to the catalytic activity center of Senecavirus (SVA) 3C protease. An in-vitro FRET enzyme activity experiment proves that the inhibition rate of 50 [mu] M of anethomycin on SVA 3C protease reaches up to 88.5%. Cellular level experiments show that the anethomycin can significantly inhibit replication of SVA viruses, shows broad-spectrum antiviral activity on various viruses such as encephalomyocarditis viruses (EMCV), porcine reproductive and respiratory syndrome viruses (PRRSV) and porcine deltacoronaviruses (PDCoV), and is low in cytotoxicity and high in selectivity index (SI).

The invention belongs to the technical field of medicines, and provides a method for screening a mutant strain and a high-virulence Seneca virus A mutant strain. The method for promoting Senecavirus A mutation provided by the invention comprises the following steps: inserting an RNAi target sequence between 3Dpol and a 3'untranslated region of an SVA starting strain; the target sequence of the RNAi contains a complementary sequence of an endogenous sequence of the miRNA. According to the research, mutagenesis is carried out on SVA by constructing a high RNAi pressure system, and a method for screening the mutant strain SVA is carried out by simulating an antiviral RNAi mechanism in a host, so that a natural evolution process of the virus is better met, a large number of stably inherited mutant strains can be obtained, and an efficient research and development means is provided for vaccine research and development aiming at the virus.

The invention belongs to the field of biology, and particularly relates to construction of a recombinant Seneca virus strain with Seneca virus 2B protein translation block site mutation. The 26th lysine (K) of the 2B protein is found to be a key site for inhibiting the translation of the host protein for the first time, and after the 26th K of the 2B protein is mutated into the alanine (A), the translation retardation phenomenon of the host protein is obviously weakened; secondly, the 26th K of the 2B protein is mutated into A to construct infectious clone, and the recombinant Seneca virus strain rSVA-2B K26A with 2B protein translation retardation site mutation is successfully rescued and obtained, compared with wild type SVA, after the rSVA-2B K26A infects host cells, the translation retardation phenomenon of the host cells is weakened, the expression level of host antiviral inflammatory cytokines is enhanced, and the expression level of the recombinant Seneca virus strain rSVA-2B K26A is improved. The Seneca virus vaccine strain can be used as a potential vaccine candidate strain, and a foundation is laid for developing a novel prevention and control strategy aiming at the Seneca virus.

The utility model discloses a kind of Seneca virus differentiation diagnosis kits of distinguishable vaccine immunization and natural infection, including diagnostic reagent kit, sample adding unit, the sample adding unit is placed on the upper portion of diagnostic reagent kit, the sample adding unit includes syringe and push rod, for pushing push rod to inside detection reagent inside syringe inside diagnostic reagent kit Synchronous quantitative push, protection unit, the protection unit is placed below sample adding unit, the protection unit includes sleeve and sliding pipe, under the sleeve and sliding pipe and the bottom sleeve of syringe limit, can avoid foreign matter into syringe and diagnostic reagent kit inside, influence reagent solution diagnostic precision, under the limit of sample adding unit, it can be added to the same amount of reagent solution inside diagnostic reagent kit simultaneously, make diagnostic reagent kit diagnostic condition same, under the limit of protection unit, it can be protected to the sample adding place of syringe and diagnostic reagent kit, avoid foreign matter adhere on reagent solution, influence detection precision.

The utility model relates to the technical field of chemiluminescence detection kit, specifically to the chemiluminescence detection kit of Seneca virus VP1 protein monoclonal antibody, including protection subassembly, the inside of the placing box is provided with clamping assembly, the clamping assembly includes the installation strip, the installation strip side surface is provided with sliding slot, the sliding slot quantity is two and the position corresponds, the sliding groove inner wall sliding connection has the slide, the slide quantity is ten and the adjacent two slides are a group, and the fixed connection has the spring between every group two slides, the slide side surface fixed connection has semicircle clamping plate, the semicircle clamping plate quantity is several and the position corresponds to adjacent two semicircle clamping plates, the installation strip side surface fixed connection is in the placing box inner wall, through the elastic clamping adaptation different specifications sampling tube, prevent it from shaking and falling off in the operation, reduce sample spilling, the protection box body can also block spilled sample when accidentally, guarantee detection.

The present application relates to the technical field of biotechnology, and particularly relates to the use of compound GC376 in preventing or treating Senecavirus disease. The compound GC376 is added in preparation or medicine to inhibit Senecavirus replication. The method realizes effective inhibition of Senecavirus replication in host cells by using the compound GC376. Meanwhile, the present application also provides a medicine combination for inhibiting Senecavirus replication. It is verified by experiments that the compound GC376 can significantly reduce the titer of Senecavirus, reduce the expression of viral proteins, inhibit the replication of Senecavirus, and has a good inhibitory effect on Senecavirus infection, thereby providing a new way and strategy for the prevention and treatment of Senecavirus disease.

The invention belongs to the technical field of PCR (polymerase chain reaction) detection, and particularly relates to a method, a primer group and a kit for simultaneously detecting African swine fever virus, Seneca virus and porcine circovirus type 2 on the basis of triple PCR. DNA of a sample to be detected is extracted, a triple PCR reaction system is prepared, the DNA of the sample to be detected is used as a template, real-time fluorescent quantitative PCR amplification is performed by using the primer group, and the kit is used for simultaneously detecting the African swine fever virus, the Seneca virus and the porcine circovirus type 2. When Ct is less than or equal to 35 and a specific amplification curve appears, the result is positive; when Ct is larger than 35 and smaller than 40, repeated detection should be carried out, and if the result is still that Ct is larger than 35 and smaller than 40 and a specific amplification curve appears, the result is positive; when Ct is greater than or equal to 40 or Ct cannot be detected, the result is negative. The method is fast, low in cost, simple to operate, low in signal interference during synchronous detection, high in efficiency and high in accuracy; and the kit also has high efficiency, specificity and high sensitivity.

The application belongs to the technical field of biology, and discloses a preparation method of a Salmonella typhimurium flagellin and Senecavirus antigen fusion protein. The application selects Escherichia coli Rosetta (DE3) as a host, fuses a Salmonella typhimurium flagellin gene and a Senecavirus antigen gene, optimizes codons, and then clones the fusion protein into a pET-28a (+) vector, so that the obtained fusion protein can keep the adjuvant activity of the flagellin, improve the solubility and expression level of VP2, and the yield can reach 40 mg / L, thereby greatly reducing the expression and purification process cost. The fusion protein provided by the application can activate the TLR5 path and activate mouse macrophages, and the production process is simple and low in cost, so that the application has important significance for the research and production of Senecavirus subunit vaccines.

The invention is applicable to the field of genetic engineering and biotechnology detection, and provides a nano antibody specifically bound with Seneca virus A and application of the nano antibody. The nucleotide sequence of the nano antibody is as shown in SEQ ID NO: 1, and the amino acid sequence of the nano antibody is as shown in SEQ ID NO: 2. The anti-SVA nano antibody provided by the invention is relatively high in titer and good in specificity; the detection method provided by the invention is used for detecting the Senecavirus A antibody, and has good specificity and sensitivity.

The invention relates to the technical field of biology, in particular to application of TRIM38 gene / protein as a target spot to screening of drugs for preventing or treating Seneca virus and application of an expression activator or an activity enhancer of the TRIM38 gene / protein to preparation of drugs for preventing or treating Seneca virus infection. The invention proves that TRIM38 can play an anti-SVV role by inhibiting SVV virus titer and structural protein expression for the first time, and provides a new target and strategy for prevention and treatment of SVV infection.

The application discloses application of a host PRKCSH gene as a target point in prevention and treatment of various RNA virus infections. Experiments prove that knocking out the PRKCSH gene of a pig can significantly inhibit adhesion and invasion of Japanese B encephalitis virus to a host cell, and can also significantly inhibit replication of a classical swine fever virus, a porcine transmissible gastroenteritis virus, a Senecavirus, a porcine epidemic diarrhea virus and a porcine reproductive and respiratory syndrome virus in the host cell. In addition, knocking out the PRKCSH gene of a human can significantly inhibit replication of the Japanese B encephalitis virus in the host cell. Therefore, specific targeted editing of the PRKCSH gene or interference with protein expression of the PRKCSH gene can inhibit infection of various RNA viruses to host cells.

The application belongs to the technical field of biology, and discloses a Salmonella typhimurium flagellin derivative and Senecavirus antigen fusion protein and application thereof. The Salmonella typhimurium flagellin derivative gene and the Senecavirus antigen gene are fused, and after codon optimization, the fusion protein is cloned into a pET-28a(+) vector, and the fusion protein is expressed by using an Escherichia coli system. In-vivo experiment verification shows that the fusion protein has good biological activity, can activate TLR5 ligand activity, and promote macrophage activation, and lays a foundation for development of a Senecavirus subunit vaccine.

The invention relates to the technical field of biology, in particular to application of a compound GC376 to prevention or treatment of Seneca virus diseases. The compound GC376 is added into a preparation or a medicine to inhibit Seneca virus replication. The method achieves effective inhibition of senecavirus replication in host cells by using a compound GC376. Meanwhile, the invention also provides a pharmaceutical composition for inhibiting Senecavirus replication. Experiments prove that the compound GC376 can significantly reduce the titer of Seneca virus, reduce the expression of viral protein and inhibit the replication of Seneca virus, has a good inhibition effect on Seneca virus infection, and provides a new way and strategy for prevention and treatment of Seneca virus disease.

The application discloses a detection kit capable of distinguishing porcine Senecavirus infection or vaccine immunization, and belongs to the technical field of animal virus antibody detection. VP2-VP3-VP1 and 3AB-3C recombinant proteins used in the application have stronger antigenicity and diagnostic sensitivity when used for detecting porcine Senecavirus, can effectively distinguish natural infection from vaccine immunization, and thus realize accurate detection. In addition, the detection method also has good specificity, analysis sensitivity, repeatability and reproducibility, has a wider application range, and has important application value.

The application belongs to the technical field of biotechnology and relates to the field of veterinary diagnosis, and particularly discloses a liquid-phase blocking ELISA kit for porcine Senecavirus neutralizing antibody, which contains porcine Senecavirus single-domain antibody and matched detection reagents, and the porcine Senecavirus single-domain antibody can also be an antigen-binding fragment of the single-domain antibody, the single-domain antibody is selected from one or more groups in 52 groups, the heavy chain variable region of each group of single-domain antibodies or antigen-binding fragments contains CDR-1, CDR-2 and CDR-3, the amino acid sequences of the heavy chain variable region CDR-1 (VH-CDR-1) of the 52 groups of single-domain antibodies or antigen-binding fragments are respectively shown in SEQ ID NO. 1-SEQ ID NO. 52, the amino acid sequences of the heavy chain variable region CDR-2 (VH-CDR-2) are respectively shown in SEQ ID NO. 53-SEQ ID NO. 104, and the amino acid sequences of the heavy chain variable region CDR-3 (VH-CDR-3) are respectively shown in SEQ ID NO. 105-SEQ ID NO. 156. The kit can quickly detect the SVA neutralizing antibody level in serum, has high sensitivity, good specificity and stable results, and can be better applied to SVA vaccine efficacy evaluation.

The invention discloses a method for rapidly quantifying Seneca virus inactivated antigen protein. The method comprises the following steps: establishing a specific purification technology to obtain a Seneca antigen standard substance, and establishing a linear relation equation between the Seneca antigen content and the chromatographic peak area; meanwhile, a pretreatment method for samples which are not suitable for direct sample loading detection in the Seneca inactivated vaccine production process is established, so that the samples which can be used for detection are obtained; further, in order to eliminate the influence of the target protein loss on the detection result in the sample treatment process, determining the sample recovery rate R in different treatment processes through a labeling recovery test; and finally, through a linear equation and the recovery rate R, calculating to obtain the antigen content of the sample at each stage in the Seneca inactivated vaccine production process. The method is used for directly detecting the content of the Seneca inactivated antigen protein, has the advantages of simple detection operation process and high detection result repeatability, is suitable for each stage of the Seneca inactivated vaccine production process, and can effectively control the Seneca inactivated vaccine production process and the vaccine efficacy.

The invention belongs to the technical field of medicines, and particularly relates to application of kaempferide in preparation of anti-Seneca virus medicines. The invention also discloses an anti-Seneca virus pharmaceutical composition of the kaempferide. Experiments prove that the kaempferide can obviously inhibit the replication of Seneca virus in both BHK-21 cell and PK-15 cell models, and has no obvious toxicity to cells in an effective action concentration range. Through deep study on the antiviral mechanism, the kaempferide can enhance the antiviral reaction by regulating the innate immune pathway of host cells, and a new treatment strategy and drug choice are provided for infectious diseases caused by Seneca virus.

This invention discloses a porcine Fab antibody against Seneca virus A (SVA) and its competitive ELISA detection method. The amino acid sequences of the heavy and light chain variable regions of the neutralizing Fab genetically engineered antibody are shown in SEQ ID No. 4 and SEQ ID No. 7. The antibody exhibits high affinity for SVA. A competitive ELISA detection method was established, determining the optimal SVA particle coating concentration to be 1 μg / mL, the optimal working concentration of the competitive antibody to be 0.5 μg / mL, the optimal dilution ratio of the serum sample to be tested to be 1:10, and the optimal dilution ratio of the enzyme-labeled avidin to be 1:30000. The highest sensitivity and specificity (96.88% and 100%, respectively) were achieved when the inhibition percentage (PI) was 47%, and the antibody did not react with positive sera from common major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, and the inter-assay coefficient of variation ranged from 1.19% to 11.72%, indicating good reproducibility. This method and VNT detection were used to simultaneously detect 224 clinical porcine serum samples, with a concordance rate of 93.75%.

The application provides a kind of chimeric flagellin and its application.The chimeric flagellin is formed by the chimeric of A type Seneca virus VP2 fragment and Escherichia coli Nissle 1917 flagellin FliC through Linker (GGGGS) 3, which realizes the soluble expression of chimeric flagellin in Escherichia coli system, and the yield can reach 200 mg / L at most. And through the detection of in vitro Caco2 and Raw264.7 cell models, the recombinant chimeric flagellin shows good TLR5 receptor activity, which can effectively activate TLR5 signal pathway.

The application belongs to the technical field of medicine, and particularly relates to application of phyllangin in preparation of anti-senecavirus virus (SVV) drugs. The phyllangin is separated from seeds of phyllanthus emblica, and the purity of the phyllangin reaches more than 99%. The effective concentration of the phyllangin in the anti-SVV drug is in the range of 1-20 muM. The application further provides a preparation for treating SVV, and the preparation comprises an active ingredient of phyllangin. The application firstly finds that the phyllangin has the effect of resisting SVV, the phyllangin as a natural product has low cytotoxicity to host cells, has small toxic and side effects, and has a relatively wide source, and the application of the phyllangin to preparation of anti-SVV drugs provides a new choice for treatment of SVV infection.

The present application relates to the technical field of molecular biology, and particularly relates to a type A Seneca mutant virus strain and application. The present application obtains a Seneca mutant virus strain CRL-FJ-P80 by continuously subculturing a type A Seneca virus CH-FJ-2017 strain in CRL-2843 cells for 80 generations. Compared with the CH-FJ-2017 parent strain, the virus nucleic acid and virion replication levels of the Seneca mutant virus strain CRL-FJ-P80 are both significantly increased, indicating that the Seneca mutant virus strain CRL-FJ-P80 has stronger replication and production capacity. Moreover, the antigenicity of the Seneca mutant virus strain CRL-FJ-P80 does not change, and the immune capacity is enhanced. The Seneca mutant virus strain CRL-FJ-P80 can be used as a candidate strain of inactivated vaccine, and is used for preparation of Seneca virus inactivated vaccine, and has a wide application prospect.

The invention belongs to the technical field of medicines, and particularly relates to an application of karanjin in preparation of anti-Seneca virus medicines. The karanjin is separated from seeds of pongamia pinnata, and the purity of the karanjin reaches 99% or above; the effective concentration range of the karanjin in the Seneca virus resisting medicine is 1 to 20 [mu] M. The invention also provides a product for treating Seneca virus. The product comprises the active ingredient, namely the karanjin. It is found for the first time that the karanjin has the anti-Seneca virus effect, the karanjin serves as a natural product, toxicity to host cells is low, toxic and side effects are small, the source is relatively wide, the karanjin is applied to preparation of the anti-SVV medicine, and a new choice is provided for treatment of SVV infection.

The application provides an application of harringtonine in preparation of an anti-Senecavirus preparation and belongs to the technical field of preparation of antiviral drugs. The application provides a new use of harringtonine, the harringtonine can effectively inhibit replication of Senecavirus and reduce activity of the Senecavirus, and can be used for preparation of an anti-Senecavirus preparation.

The present disclosure provides immunogenic compositions and vaccines as well as associated methods for control of Senecavirus A (SVA) in pigs. Advantageously, the compositions and methods described herein can be administered using a single dose and can be delivered to pigs via different routes. The compositions and methods utilize a live attenuated vaccine strategy that provides robust efficacy and near sterilizing immunity in pigs.

The application discloses a group of antigen polypeptides of porcine Seneca Valley virus and application thereof; the antigen polypeptide of the porcine Seneca Valley virus comprises a fragment shown in sequence SEQ ID NO.1, or a sequence obtained by substituting, deleting or adding one or more amino acids to sequence SEQ ID NO.1 and having immunogenicity. Compared with a porcine Seneca Valley epitope vaccine product under research, the antigen polypeptide of the porcine Seneca Valley virus and application thereof in the field of vaccines have the following advantages: the application can stabilize the spatial conformation of the antigen polypeptide by limiting the polypeptide epitope, and can significantly improve the immunogenicity of the original antigen polypeptide.

This invention belongs to the field of molecular biology, specifically relating to the application of SND1-overexpressing cells in promoting small RNA virus replication. The invention first constructs an SND1 gene overexpression vector, transfects it into BHK-21 cells, and then selects a stable cell line. Infection of the cell line with foot-and-mouth disease virus (FMDV) and Seneca virus (SVV) revealed that the titers of FMDV and SVV, as well as the expression levels of viral RNA and proteins, were significantly higher than in wild-type cells, indicating that SND1 promotes FMDV and SVV replication. The constructed SND1-overexpressing cells can efficiently proliferate FMDV and SVV and can be used as cell lines for FMDV and Seneca virus vaccine production. This provides a new tool for FMDV and SVV mechanism research, vaccine preparation, and the development of detection reagents, and has significant scientific research and application value.

Use of a CRYZL1 gene / protein expression inhibitor or knockout agent in the preparation of a drug for preventing or treating a Seneca virus infection. The drug can inhibit the endocytosis, growth, proliferation, and invasion of the Seneca virus. A pharmaceutical combination for preventing or treating a Seneca virus infection, comprising a CRYZL1 gene / protein expression inhibitor or knockout agent. Use of the CRYZL1 gene in the preparation of a formulation for breeding a Seneca virus-resistant breed of pig or in the preparation of a formulation for improving the germplasm resource of a breed of pig with Seneca virus-related resistance.

The invention belongs to the technical field of medicines, and provides a Seneca virus A mutant strain and application thereof. The insertion mutation of the screened mutant strain SVA genome at the 1494 site amino acid changes the spatial conformation of the virus 3B protein, enhances the virus replication efficiency and interferes the host immune response; the mutant strain has high titer and hereditary stability, and can solve the problem of virus dosage fluctuation in a traditional challenge experiment; the genome stability and high replication efficiency of the strain provide a standardized tool for vaccine research and development.

The invention relates to the technical field of genetic engineering, in particular to Encapsulin fusion protein for expressing Seneca virus epitope and application of Encapsulin fusion protein in preparation of subunit vaccines. The invention discovers that Seneca virus antigen epitopes are inserted among the 62nd to 63rd positions, the 124 to 125th positions, the 138 to 139th positions and the 239th to 240th positions of amino acids of an Encapsulin protein fragment subjected to amino acid sequence modification, and / or the 62nd to 63rd positions, the 124 to 125th positions, the 138 to 139th positions and the 239th to 240th positions of amino acids are replaced by the Seneca virus antigen epitopes, so that efficient and soluble expression of a target antigen in escherichia coli can be realized; the protein cage nano antigen particles are successfully self-assembled; the protein nano antigen particle can induce a widely neutralizing Seneca virus antibody, improves the immune efficacy, and has the potential of becoming a Seneca multi-epitope vaccine.