65 results about "Senecavirus" patented technology

The present invention discloses a porcine Seneca virus neutralization liquid phase blocking ELISA kit, which comprises an effective amount of porcine Seneca virus antigen, an effective amount of porcine Seneca virus single domain antibody 1, an effective amount of porcine Seneca virus single domain antibody 2, and a matching detection reagent. The porcine Seneca virus antigen is porcine Seneca virus particles purified by sucrose density gradient centrifugation, and the particle size of the purified porcine Seneca virus particles is 20 to 30 nm. The amino acid sequence of the heavy chain variable region of the porcine Seneca virus single domain antibody 1 is shown in SEQ ID NO.1. The amino acid sequence of the heavy chain variable region of the porcine Seneca virus single domain antibody 2 is shown in SEQ ID NO.2. The kit of the present invention has the advantages of high efficiency, simplicity, sensitivity, stability, etc., has a good correlation with the traditional diagnostic method virus neutralization experiment, and is suitable for a wide range of applications.

The invention belongs to the technical field of biology, and discloses a preparation method of fusion protein of salmonella typhimurium flagellin and Seneca virus antigen. Escherichia coli Rosetta (DE3) is selected as a host, a salmonella typhimurium flagellin gene and a Seneca virus antigen gene are fused and are cloned to a pET-28a (+) vector after codon optimization, the obtained fusion protein keeps the flagellin adjuvant activity, the solubility and the expression level of VP2 are improved, the yield can reach 40 mg / L, and the fusion protein can be used for preparing a recombinant vector. The expression and purification process cost is greatly reduced. The fusion protein provided by the invention can activate a TLR5 pathway and mouse macrophages, is simple in production process and low in cost, and has important significance on research and production of Seneca virus subunit vaccines.

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 provides a novel chimeric flagellin and application thereof. The novel chimeric flagellin is formed by chimeric combination of a Senecavirus A VP2 fragment and an escherichia coli Nissle 1917 flagellin FliC through Linker (GGGGS) 3, soluble expression of the novel chimeric flagellin in an escherichia coli system is achieved, and the highest yield can reach 200 mg / L. In addition, through detection of in-vitro Caco2 and Raw264.7 cell models, the recombinant novel chimeric flagellin shows good TLR5 receptor activity, and can effectively activate a TLR5 signal channel.

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 gene engineering, and particularly relates to application of an RSAD1 gene / protein serving as a target spot in screening of drugs for inhibiting picornaviridae virus infection. The silent RSAD1 gene is accidentally found to be capable of remarkably inhibiting the replication of the Seneca virus and can be used as a target spot for researching and developing an antiviral drug for the Seneca virus. The invention provides an sgRNA sequence of a targeted RSAD1 gene, a CRISPR-Cas9 gene editing technology is combined, the RSAD1 gene is silenced, an RSAD1 gene knockout monoclonal cell line is obtained, the cell line can remarkably inhibit the replication of Seneca virus, and a target spot is provided for further screening drugs and reagents for resisting the replication of Seneca virus; meanwhile, it is found that overexpression of the RSAD1 protein in host cells can significantly promote replication of Seneca viruses, and the RSAD1 protein can be used as a production cell line of Seneca viruses or vaccines and used for production of Seneca viruses or vaccines.

The invention provides a multiple nucleic acid detection technology based on CRISPR / Gs12 system mediation and application, different Gs12 nuclease are respectively combined with corresponding specific crRNA and single-stranded DNA reporter genes, and then a recombinase polymerase amplification (RPA) method is combined, so that quadruple, triple or double specific accurate detection of target nucleic acid molecules can be realized. Therefore, a nucleic acid detection technology and a kit capable of simultaneously distinguishing the Seneca virus (SVA), the hog cholera virus (CSFV) and the African hog cholera virus (ASFV) are developed. The method provided by the invention can realize rapid multiple detection of target nucleic acid, and has good popularization and application values in the field of nucleic acid detection.

The invention provides application of dipotassium glycyrrhizinate in preparation of a preparation for resisting porcine Seneca virus, and further provides a product for preventing and / or treating SVA infection. The invention provides the application of dipotassium glycyrrhizinate in preparation of the preparation for preventing and / or treating SVA infection, the dipotassium glycyrrhizinate has a very obvious anti-SVA infection effect, cell infection caused by SVA can be reduced, the adsorption, cell entry and assembly stages of SVA can be inhibited, and a scientific and reliable theoretical basis is provided for clinical treatment of the porcine Seneca virus disease.

The invention relates to the technical field of molecular biology, in particular to an A-type Seneca mutant virus strain and application thereof. According to the invention, a Seneca virus CH-FJ-2017 strain A is subjected to continuous subculture for 80 generations in a CRL-2843 cell, and then a Seneca mutant virus strain CRL-FJ-P80 is obtained; compared with a CH-FJ-2017 parent virus strain, the Seneca mutant virus strain CRL-FJ-P80 disclosed by the invention has the advantages that the virus nucleic acid and virion replication levels are obviously increased, so that the Seneca mutant virus strain CRL-FJ-P80 disclosed by the invention has stronger replication and production capacity; the antigenicity of the Seneca mutant virus strain CRL-FJ-P80 is not changed, and the immunocompetence of the Seneca mutant virus strain CRL-FJ-P80 is enhanced; the strain can be used as an inactivated vaccine candidate strain, is used for preparing Seneca virus inactivated vaccines, and has a wide application prospect.

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 present invention relates to the field of molecular biology technology, and specifically to an application of a molecular chaperone TF in promoting the soluble expression of type A Seneca virus structural protein VP2. The present invention first constructs a SVA VP2 protein recombinant expression plasmid pET28a-SVA-VP2 based on the gene sequence of the SVA CH / FJ / 2017 strain; then the pET28a-SVA-VP2 recombinant plasmid and the molecular chaperone pTf16 plasmid are co-transfected into BL21 (DE3) competent cells to successfully prepare a soluble recombinant SVA VP2 protein; the recombinant VP2 protein has good immunogenicity in both in vivo and in vitro tests, and can induce the body to produce a strong humoral immune response and cellular immune response, providing experimental materials and theoretical basis for the development of SVA subunit vaccines and diagnostic preparations.

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.

This invention belongs to the field of genetic engineering, specifically relating to the application of RSAD1 gene / protein as a target in screening drugs to inhibit infection by microribonucleoviridae viruses. This invention unexpectedly discovered that silencing the RSAD1 gene can significantly inhibit Seneca virus replication, and can be used as a target for the development of antiviral drugs against Seneca virus. This invention provides an sgRNA sequence targeting the RSAD1 gene, and combined with CRISPR-Cas9 gene editing technology, silences the RSAD1 gene, obtaining an RSAD1 gene knockout monoclonal cell line. This cell line can significantly inhibit Seneca virus replication, providing a target for further screening of drugs and reagents to resist Seneca virus replication. Simultaneously, this invention found that overexpression of the RSAD1 protein in host cells can significantly promote Seneca virus replication, and can be used as a production cell line for Seneca virus or vaccines.

The present invention provides a use of Corydalis for preparing a product for preventing and / or treating SVA infection; in another aspect, the present invention also provides a product for preventing and / or treating SVA infection, wherein the product contains Corydalis at a pharmacologically effective concentration. The present invention provides the use of Corydalis in preparing a preparation for preventing and / or treating SVA infection. Corydalis has a very significant effect on preventing SVA infection, can reduce cell infection caused by SVA, and inhibit the adsorption, endocytosis, replication, assembly and release stages of SVA, providing a scientific and reliable theoretical basis for the clinical treatment of swine Seneca virus disease.

The invention belongs to the technical field of preparation of veterinary vaccines, and particularly relates to a Senecavirus type A nanoparticle antigen and application thereof in preparation of a nanoparticle vaccine. According to the invention, a general Th cell epitope is fused with a highly conservative B cell neutralizing epitope VP2150-160aa of a three-copy Senecavirus A capsid protein, and the fused recombinant protein is displayed on the surface of a nanoparticle, so that the Senecavirus A nanoparticle antigen is finally constructed. The Senecavirus A nanoparticle vaccine is prepared from the Senecavirus A nanoparticle antigen, and the Senecavirus A nanoparticle vaccine can display more antigens at a time, so that the immunogenicity of the vaccine can be improved, and the immune effect of the vaccine can be enhanced.

This invention belongs to the field of biotechnology, specifically relating to a strain of Bacillus licheniformis, an antibacterial agent, a method for preparing antibacterial active substances, and their applications. The Bacillus licheniformis DCGGN-121 provided by this invention has the preservation number CCTCC NO: M 2024878. The bacteriocins prepared from this strain are effective against African swine fever virus (ASFV), Seneca virus (SVA), and enterovirus 71 (EV71). The results of the examples show that the bacteriocins prepared from Bacillus licheniformis DCGGN-121 can significantly reduce the replication levels of ASFV, SVA, and EV71, as well as reduce the expression of ASFV-p72, SVA-VP2, and EV71-VP1 proteins. Bacillus licheniformis DCGGN-121 can be used to prepare natural microecological preparations, suitable for livestock farming.

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 discloses a Seneca virus nucleic acid detection composition and a Seneca virus nucleic acid detection kit based on RT-RPA and CRISPR-Cas13a and application of the Seneca virus nucleic acid detection composition and the Seneca virus nucleic acid detection kit. The composition comprises an RT-RPA (reverse transcription-recombinase polymerase amplification) constant-temperature amplification system and a CRISPR-Cas13a (clustered regularly interspaced short palindromic repeats The RT-RPA isothermal amplification system comprises an upstream primer and a downstream primer; wherein the nucleotide sequence of the upstream primer is as shown in SEQ ID NO. 16; the nucleotide sequence of the downstream primer is as shown in SEQ ID NO. 17; the CRISPR-Cas13a detection system comprises specific crRNA (Complementary Ribonucleic Acid) of a targeted Senecavirus, LwaCas13a protein and RNase report, wherein the nucleotide sequence of the specific crRNA is as shown in SEQ ID NO.18. The CRISPR-Cas13a detection system is used for detecting the Senecavirus, the LwaCas13a protein and the RNase report, and the CRISPR-Cas13a detection system is used for detecting the Senecavirus. Compared with a real-time fluorescent quantitative RT-PCR (qRT-PCR) detection method, the method established by the invention has the advantages that the detection performance is equivalent to that of the qRT-PCR, but compared with the qRT-PCR, the detection speed is higher, the operation procedures are fewer, and the method is more suitable for in-situ detection.

The invention provides application of harringtonine in preparation of anti-Seneca virus preparations, and belongs to the technical field of preparation of antiviral drugs. The invention provides new application of harringtonine, and harringtonine can effectively inhibit replication of Seneca virus and reduce activity of Seneca virus, and can be used for preparing anti-Seneca virus preparations.

The present invention relates to killed / inactivated and / or recombinant Senecavirus A immunogenic compositions and vaccines, and methods of using such immunogenic compositions and vaccines to prevent or treat animals in need thereof.

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 invention belongs to the technical field of biology, and particularly relates to bacillus licheniformis, a bacteriostatic agent, a method for preparing bacteriostatic active substances and application. The preservation number of the bacillus licheniformis DCGGN-121 provided by the invention is CCTCC (China Center For Type Culture Collection) NO: M 2024878. The bacteriocin prepared from the strain can resist African swine fever virus (ASFV), Seneca virus (SVA) and enteritis virus 71 (EV71 virus). The results of the embodiment show that the bacteriocin prepared from the bacillus licheniformis DCGGN-121 can obviously reduce the replication level of ASFV (Allergic Sequence Factor Virus), SVA (Seaweed Vector Amplification) and EV-71, and reduce the expression of ASFV-p72, SVA-VP2 and EV71-VP1 proteins. The bacillus licheniformis DCGGN-121 can be used for preparing a natural microecological preparation and is suitable for the food industry and the livestock breeding industry.

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.