261 results about "Classical swine fever" patented technology

The invention relates to the technical field of biology, and aims to provide a monoclonal antibody 7A12 for identifying gene type 2 swine fever as well as a preparation method and an application method of the monoclonal antibody 7A12. The monoclonal antibody comprises Ig structural domains VH CDR1, VH CDR2 and VH CDR3 of a heavy chain of the antibody, and Ig structural domains VL CDR1, VL CDR2 and VL CDR3 of a light chain of the antibody, wherein the amino acid sequences of the VH CDR1, the VH CDR2 and the VH CDR3 are respectively as shown in SEQ ID NO: 1 to SEQ ID NO: 3; the amino acid sequences of the VL CDR1, the VL CDR2 and the VL CDR3 are respectively as shown in SEQ ID NO: 4 to SEQ ID NO: 6. The antibody provided by the invention has the advantages of high titer, strong specificity, high affinity and the like; the kit has the advantages of high sensitivity and strong specificity, has no cross reaction with the type 1 swine fever vaccine strain, and can identify and diagnose swine fever gene type 2 wild strain infection and vaccine immunity in serology; the method can be further used for establishing a blocking ELISA kit for identifying the gene type 2 hog cholera, and can effectively diagnose and purify the gene type 2 hog cholera virus.

The invention discloses an application of phloretin in preparation of a medicine or a feed additive for treating or preventing swine envelope virus infection, the medicine is phloretin or oral liquid, granules, tablets and powder prepared from phloretin as an effective component and a solvent or an auxiliary material, and the use dosage is 2.5-10mg / kg of the body weight of a pig; or the phloretin is directly added into the feed, and the addition amount of the phloretin is 0.01-0.03% of the total weight of the pig feed. The phloretin or the composition containing the phloretin is used as a medicine or a feed additive, is high in safety and free of toxic and side effects, and has the effects of remarkably inhibiting replication of envelope viruses such as porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus and African swine fever virus and reducing the toxicity of the porcine envelope viruses; the defects of immune failure possibly caused by vaccination, virus variation caused by use of antiviral drugs and the like are overcome, the morbidity and mortality of the piglets infected with the swine envelope virus are effectively reduced, and the effect of preventing and treating or preventing the swine envelope virus infection diseases is achieved.

An immunogenic fragment of African swine fever virus CD2v protein, a recombinant protein, an immunogenic composition, and a use thereof. The provided immunogenic fragment of African swine fever virus CD2v protein or a variant thereof with immunogenicity can greatly improve the expression quantity while retaining the strong immunocompetence.

The invention discloses a fluorescent PCR (polymerase chain reaction) detection reagent and kit for detecting African swine fever virus and application of the fluorescent PCR detection reagent and kit. The fluorescent PCR detection reagent comprises a primer probe group, in the primer probe group, the sequences of primers are shown as SEQ ID NO: 1 and SEQ ID NO: 2, and the sequence of probes is shown as SEQ ID NO: 3; the fluorescent PCR detection reagent for detecting the African swine fever virus, provided by the invention, has the advantages of good anti-interference effect, high sensitivity and good specificity, and can be effectively used for detecting the African swine fever virus.

The invention discloses a method for simulating a spreading rule of African swine fever virus aerosol in a pig farm, and relates to the technical field of animal epidemic disease spreading simulation. Comprising the following steps: selecting fluorescent microspheres, and preparing a fluorescent microsphere suspension; building a fluorescent microsphere suspension release system; dividing a pig farm into a plurality of cells, and arranging a sampling network in the pig farm; starting all the samplers of the sampling network, idling for a certain time, and closing all the samplers after measuring the environment background concentration; synchronously starting a fluorescent microsphere suspension release system and samplers of all sampling networks, and synchronously recording environment temperature and humidity, wind speed and wind direction; and performing quantitative analysis on the fluorescence of the pig farm at different time points, and generating a distribution diagram and a dynamic change diagram of the fluorescence in the pig farm according to an analysis result. According to the invention, by releasing the fluorescent microspheres with matched particle sizes and combining with a gridding monitoring network, a distribution diagram and a dynamic change diagram during virus diffusion are dynamically generated, and data support is provided for pig farm ventilation optimization and accurate disinfection.

The invention provides application of thalictriine in preparation of antiviral drugs, and belongs to the technical field of antiviral drugs. The invention proves that the thalictriine has an inhibition effect on various common infectious disease viruses of domestic pigs from a cellular level, and particularly has a remarkable inhibition effect on proliferation of African swine fever virus (ASFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2) and porcine epidemic diarrhea virus (PEDV) in vitro. The invention also discloses an antiviral composition consisting of the active ingredient thalictriine or the pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier and / or excipient, and discloses an application of the composition in preparation of a virus infection inhibitor and / or a pharmaceutical preparation for preventing and / or treating virus infectious diseases. And a basis is provided for developing novel broad-spectrum antiviral drugs.

The present invention discloses an application of a nano-luciferase fusion antigen in the detection of African swine fever virus antibodies. The fusion antigen is composed of LgBiT-p30 and SmBiT-p30 proteins, which are fused and expressed by two subunits of nano-luciferase, LgBiT and SmBiT, respectively, with the African swine fever virus p30 protein. The fusion antigen can bind to a specific antibody against the African swine fever virus p30 protein to form a functional luciferase, which reacts with a substrate to produce a bright luminescent signal. The fusion antigen has high specificity and sensitivity in the detection of African swine fever virus antibodies, making the antibody detection process more efficient and rapid. The present invention realizes simple, rapid, sensitive and specific detection of African swine fever virus antibodies, and provides an innovative and reliable solution for the serological diagnosis of African swine fever.

The invention discloses an African swine fever virus T cell epitope peptide for inducing swine T cell immune response and a composition of the African swine fever virus T cell epitope peptide. The amino acid sequence of the African swine fever virus T cell epitope peptide is shown as any one of SEQ ID NO.1-19. Or an amino acid sequence which is derived from the amino acid sequence shown in SEQ ID NO.1-19 and maintains the protein function shown in SEQ ID NO.1-19 through substitution and / or deletion and / or addition of one or more amino acid residues of the amino acid sequence shown in SEQ ID NO.1-19; the composition comprises amino acids of which the sequences are SEQ ID NO.1-19. A group of ASFV key T cell antigens and epitope peptides are obtained by combining IFN-gamma ELISpot and flow cytometry screening, the ASFV key T cell antigens and epitope peptides can be used as vaccine components to induce strong T cell immune response, and T cells activated by the antigen peptides have the maximum immune protection potential and are essential key antigens for ASFV vaccine research and development.

Provided are an immunogenic fragment, a recombinant protein, and an immunogenic composition of the African swine fever virus E248R protein, as well as the use thereof. The immunogenic fragment of the African swine fever virus E248R protein or an immunogenic variant thereof can greatly improve expression levels while retaining strong immunological activity.

The invention belongs to the technical field of molecular markers, and particularly relates to an SNP (Single Nucleotide Polymorphism) molecular marker of a PRRX2 gene associated with an African swine fever virus antibody level, a primer and application. The locus of the SNP molecular marker is located at the 143bp position of the third exon of the porcine PRRX2 gene with the gene version number of XM005654574.2, the SNP molecular marker has T / C polymorphism, and the nucleotide sequence of the SNP molecular marker is shown as SEQ ID NO.3. The invention also discloses a kit for detecting the porcine PRRX2 gene with the gene version number of XM005654574.2. The molecular marker provided by the invention is closely associated with the African swine fever virus antibody level of the pig, and compared with the pig with the genotype of TC / CC at the site, the pig blood with the genotype of TT has a higher African swine fever virus antibody level; the molecular marker can be applied to breeding of high-quality breeding pigs resistant to African swine fever, so that the economic benefit of breeding pigs is effectively improved.

The invention discloses a portable immunochromatography device for detecting an African swine fever virus neutralizing antibody and application thereof.The device comprises a reading module, a sample preparation module and a detection module, the sample preparation module and the detection module are integrally connected up and down, and the sample preparation module is sequentially provided with a sample adding hole, a filtering chromatography cavity and a communicating pipe which are communicated from top to bottom; a filtering chromatography part is arranged in the filtering chromatography cavity, the detection module is provided with a fluorescence immunochromatography test strip and a shell, the fluorescence immunochromatography test strip is located in the shell, a sample pad, a fluorescence marker pad, an NC membrane and a water absorption pad of the fluorescence immunochromatography test strip are sequentially overlapped and adhered to a PVC bottom plate, and the two ends of a conduction pipe are communicated with the filtering chromatography cavity and the shell respectively. One end of the fluorescence immunochromatography test strip is located below the filtration chromatography component, the other end of the fluorescence immunochromatography test strip is located above the sample pad, the water absorption pad of the fluorescence immunochromatography test strip and the NC membrane are sequentially inserted into the reading module, and the reading module is connected with the shell through a buckle and reads a quality control line and a detection line of the inserted NC membrane.

The invention provides a nucleic acid probe, a primer group, a kit, a detection method and application, and relates to the technical field of biology. The nucleotide sequence of the nucleic acid probe is as shown in SEQ ID NO. 1; wherein the 3'end of the nucleic acid probe is modified with MGB, and locked nucleic acid is modified at a base position corresponding to the difference between the swine fever wild strain and the swine fever vaccine strain in the nucleotide sequence. According to the nucleic acid probe, a specific sequence is utilized to precisely target a swine fever wild strain, and locked nucleic acid modification is introduced at a differential site, so that the recognition capability on single-base mismatch is remarkably enhanced, non-specific binding to a vaccine strain is thoroughly blocked, and false positive interference is eliminated; and 3 '-terminal MGB modification is matched to improve the binding stability and sensitivity, so that the efficient and accurate identification of the swine fever wild strain under the immune background is ensured.

The invention relates to the technical field of animal epidemic disease detection, in particular to a portable African swine fever detection method based on an ERA-CRISPR / Cas12a technology, space-time separation of ERA constant-temperature nucleic acid amplification and CRISPR / Cas12a detection is realized through a specially designed double-layer reaction tube, and detection can be completed within 30 minutes at 45 DEG C by matching with a 150g portable constant-temperature nucleic acid amplification analyzer. According to the method, an African swine fever virus B646L gene is used as a target, crRNA and Cas12a protein are used for recognizing a target sequence to activate a fluorescence signal, the detection sensitivity reaches 12.1 copies / mu L, and the method has no cross reaction on PRRSV, PRV and other common swine pathogens. The instrument supports Bluetooth connection with a mobile phone to display a fluorescence curve in real time, a result can be automatically judged through a threshold value or is observed by naked eyes through ultraviolet light, the problems of expensive equipment and complicated operation in the prior art are solved, and the instrument is suitable for on-site rapid detection.

The invention discloses a pork industry chain key risk factor traceability method based on multi-kernel metric learning (MKLM for short), and relates to a pork industry chain key risk factor traceability method based on multi-kernel metric learning (MKLM for short). According to the method, heterogeneous data fusion is realized through multi-kernel metric learning (MKLM for short) according to the propagation characteristics of African swine fever biological risk factors in a circulation chain. On a pork circulation chain, each link relates to various types of data, generally including numerical data (temperature, humidity, duration and the like), classified data (whether disinfection is carried out or not, live pig varieties, hygiene grades, transportation mode types and the like) and picture data. The three types of data fusion are different from traditional data fusion, and data fusion cannot be performed by directly adopting modes of splicing, weighted average and the like. Therefore, the invention provides a multi-kernel metric learning algorithm, which comprises the following steps of: processing various types of data by using different M matrixes, processing the different types of data by using different kernel functions, mapping the data into a high-dimensional space, better capturing the relationship among the different data, fusing the data, quantifying the contribution of each link to the infection risk, and obtaining the risk of infection. And traceability is realized.

The invention discloses application of phenazine-1-carboxylic acid or medicinal salt thereof in preparation of drugs for inhibiting African swine fever virus. According to the invention, a double-report virus rASFV-Glucc / EGFP for co-expressing green fluorescent protein and Gaussian luciferase is used, and screening from 246 natural small molecule compounds finds that phenazine-1-carboxylic acid has a remarkable inhibition effect on the replication of ASFV; the inhibition effect of the PCA is further evaluated, and it is found that the half maximum cytotoxicity concentration of the PCA to primary porcine alveolar macrophages of target cells is 470.5 mu M, and the half maximum inhibition concentration of the PCA to ASFV in PAMs is 1.59 mu M. According to a dose-dependent inhibition experiment result, when the PCA concentration is 25 [mu] M, the ASFV inhibition effect can reach 100 times or more. The invention has application prospects in preparation of anti-ASFV drugs or preparations and the like.

The invention discloses a recombinant 2.1 d subtype swine fever E2 protein, a subunit vaccine and application of the subunit vaccine, and belongs to the technical field of genetic engineering. According to the 2.1 d subtype swine fever E2 protein, 344-375 amino acids of the 2.1 d subtype swine fever E2 protein are removed, and 1-343 amino acids of the 2.1 d subtype swine fever E2 protein are reserved, so that the most important antigenic epitope with a protective effect of the 2.1 d subtype swine fever E2 protein can be reserved, the space structure of the recombinant 2.1 d subtype swine fever E2 protein is basically not influenced, and the immunogenicity of the 2.1 d subtype swine fever E2 protein can be reserved to the greatest extent; in addition, the recombinant 2.1 d subtype swine fever E2 protein can be efficiently expressed in a prokaryotic expression system; furthermore, the soluble expression level of the recombinant 2.1 d subtype swine fever E2 protein can be remarkably improved through co-expression of the molecular chaperone, and after the recombinant 2.1 d subtype swine fever E2 protein is prepared into a subunit vaccine, the subunit vaccine has a good protection effect on 2.1 d subtype swine fever virus infection.

A nanobody classical swine fever virus (CSFV)-E0-Nb1 against a CSFV E0 protein, and an encoding gene and use thereof are provided, belonging to the technical field of biological detection. The nanobody CSFV-E0-Nb1 has an amino acid sequence shown in SEQ ID NO: 1 and can be expressed using an expression system. The nanobody is coupled with a quantum dot to obtain an immunochromatographic test strip for distinguishing antibodies against a CSFV E2 subunit vaccine strain from those of a wild strain infected on site, and there is a simple production process of the test strip. The immunochromatographic test strip can differentiate and diagnose the antibodies against the CSFV E2 subunit vaccine strain and the wild strain, and has the advantages of rapid, convenient, and instant detection, thus providing a new method for the detection of classical swine fever (CSF) purification.

The invention belongs to the field of biological immunity, and discloses a linear B cell epitope of African swine fever virus NP419L protein and application. The epitope sequence shows good immunoreactivity through ASFV positive pig serum recognition verification. The African swine fever positive serum can be identified by using the B cell epitope identified by the invention to carry out ELISA (Enzyme-Linked Immunosorbent Assay) experiment, and the sensitivity of the African swine fever positive serum is superior to that of NP419L protein full length and other truncated fragments. The epitope identified by the invention has high conservative property in reference ASFV strains in a plurality of popular regions in China, and has wide universality and adaptability. The kit can be widely applied to etiological diagnosis, serological diagnosis, immunological detection and disease prevention and treatment of African swine fever viruses, and can be applied to preparation of African swine fever vaccines, pathogenesis research and the like.

The invention provides a biological feed leavening agent for preventing and treating African swine fever as well as a preparation method and application of the biological feed leavening agent, and belongs to the technical field of feeds. The biological feed leavening agent disclosed by the invention is prepared from the following components in parts by weight: 5 to 25 parts of micrococcus luteus, 10 to 50 parts of Weissella mesenteroides SL7, 10 to 50 parts of lactic acid bacteria, 10 to 20 parts of bacillus licheniformis and 40 to 80 parts of clostridium butyricum. The biological feed leavening agent for preventing and treating the African swine fever, provided by the invention, is added into feed, so that the completeness of mucosal barriers of organisms can be maintained, the non-specific immunity of the organisms can be improved, and the threshold value of infectious diseases, especially the threshold value of infection of the African swine fever can be increased.

The invention provides a biological safe and efficient fattening field, and belongs to the technical field of livestock breeding. A negative pressure isolation belt is formed by the outer shell and the inner shell, so that external air is forced to flow through the visible light catalysis quantum dot layer to realize pathogen inactivation while the gas flow rate is controlled. An optical fiber sensing siphon is arranged below the slatted floor to monitor sewage pathogenic bacteria in real time. Live pigs enter the house through the third-stage buffer channel. The system breaks through the limitation of a traditional physical barrier, full-path blocking of a transmission chain of epidemic diseases such as African swine fever is achieved, and the biological safety level is improved by two orders of magnitude.

The invention discloses a primer probe combination for simultaneously detecting African swine fever virus, porcine epidemic diarrhea virus and porcine reproductive and respiratory syndrome virus, a kit and application thereof. The primer probe combination consists of a first primer probe group for detecting African swine fever virus, a second primer probe group for detecting porcine epidemic diarrhea virus and a third primer probe group for detecting porcine reproductive and respiratory syndrome virus. The primer probe combination for simultaneously detecting the African swine fever virus, the porcine epidemic diarrhea virus and the porcine reproductive and respiratory syndrome virus, provided by the invention, is high in sensitivity and good in specificity, and can be effectively used for detecting the African swine fever virus, the porcine epidemic diarrhea virus and the porcine reproductive and respiratory syndrome virus.

The application discloses a use of phenazine-1-carboxylic acid or a pharmaceutically acceptable salt thereof in preparation of a medicine for inhibiting African swine fever virus. The application uses a double reporter virus rASFV-Gluc / EGFP co-expressing green fluorescent protein and Gaussia luciferase to screen 246 natural small molecule compounds, and finds that phenazine-1-carboxylic acid has a significant inhibiting effect on replication of the ASFV; further evaluation of the inhibiting effect finds that the half maximal cytotoxicity concentration of PCA on target cells of primary porcine alveolar macrophages is 470.5 muM, and the half maximal inhibitory concentration of PCA on the ASFV in PAMs is 1.59 muM. According to a dose-dependent inhibiting experiment result, when the concentration of PCA is 25 muM, the inhibiting effect on the ASFV can reach more than 100 times. The application has application prospects in preparation of medicines or preparations against the ASFV.

The invention discloses an African swine fever virus vaccine as well as related products and application thereof, and relates to the field of vaccines. The saRNA vaccine of the African swine fever virus provided by the invention adopts an optimized antigen sequence and a saRNA skeleton, and compared with a traditional African swine fever virus vaccine, the saRNA vaccine of the African swine fever virus has the advantages of being early in antibody production period, good in safety, simple in immune procedure, higher in antibody titer and the like.

The present disclosure belongs to the field of biotechnology, and specifically relates to a seven-component antigen African swine fever subunit vaccine. The present disclosure first provides an African swine fever virus antigen protein combination composed of the African swine fever virus P34 protein, P30 protein, P54 protein, A104R protein, C129R protein, X protein, and Y protein. This African swine fever virus antigen protein combination can induce a strong immune response in the host. Furthermore, the present disclosure provides a seven-component antigen African swine fever subunit vaccine including the aforementioned African swine fever virus antigen protein combination. The seven-component antigen African swine fever subunit vaccine exhibits good immunoprotection rates against challenge with the parental virulent African swine fever virus strain, poses no biosafety risks, overcomes the difficulty that existing African swine fever subunit vaccines domestically and internationally cannot provide effective immunoprotection for pigs.

The invention discloses an African swine fever mRNA vaccine composition capable of synergistically activating and enhancing humoral immunity and cellular immunity and application of the African swine fever mRNA vaccine composition, and belongs to the technical field of biological medicines. Specifically, a plurality of key antigen proteins of the African swine fever virus are used as target spots and are connected in series through connexons to form single-chain mRNA, so that multiple immunogens can be translated from the same transcript, and multiple antibodies are induced to be generated at the same time; a conservative recombinant T cell immunogen capable of activating cellular immune response is introduced, so that the cellular immune response can be enhanced, the ADE risk can be reduced, and the cross protection capability can also be improved; in addition, a molecular adjuvant IL-12 is added into the mRNA vaccine to enhance the immune response of the African swine fever vaccine, so that stronger immune protection efficacy is induced.

The present invention belongs to a new use of verrucine, and particularly relates to the use of verrucine and verrucine extract in the preparation of antiviral drugs. Verrucine has an inhibitory effect on porcine delta coronavirus, porcine epidemic diarrhea virus, pseudorabies virus, herpes simplex virus, influenza virus or African swine fever virus. The verrucine extract and verrucine of the present invention are very effective in antiviral treatment. The verrucine extract and verrucine of the present invention are very effective in broad-spectrum antiviral treatment, are safe, have few toxic and side effects, have low drug residues and are pollution-free.

The invention discloses a recombinant antigen protein, a recombinant vector, a recombinant host cell, a virus-like particle and application thereof and a vaccine, and relates to the technical field of antigen preparation, the recombinant antigen protein comprises an icosahedral porcine circovirus capsid protein and an A104R protein loaded on the surface of the icosahedral porcine circovirus capsid protein, the A104R protein comprises a protein coded by an African swine fever virus A104R gene. The recombinant antigen protein capable of simultaneously reacting with positive serum of the African swine fever virus and positive serum of the porcine circovirus is prepared, and the vaccine prepared by mixing the recombinant antigen protein with an adjuvant can effectively stimulate a pig body to generate specific antibodies of the African swine fever virus and the porcine circovirus; therefore, reference is provided for joint defense of the two viruses. The recombinant antigen protein provided by the invention has the characteristics of stable expression, high yield and high solubility.

The invention provides an indirect ELISA antibody detection method based on African swine fever virus D250R protein, and relates to the technical field of virus detection. The method comprises the following steps: cloning a D250R protein gene into a prokaryotic expression vector pET-32a (+) to construct a recombinant plasmid pET-32a-D250R, inducing a D250R recombinant protein through IPTG (isopropyl-beta-d-thiogalactoside), purifying the recombinant protein and coating an elisa plate, and optimizing and determining the optimal conditions of indirect ELISA (Enzyme-Linked Immunosorbent Assay). On the basis of the D250R protein, serum to be detected is taken as a sample, and the established indirect ELISA antibody detection method is high in sensitivity, strong in specificity and high in coincidence rate with a commercialized kit, and provides reference for development and prevention and control of an ASF diagnosis technology.

The invention discloses a real-time detection system and a monitoring and early warning method for African swine fever virus. The system comprises a latex microsphere labeled immunochromatography antibody detection card, a portable MINI-PCR instrument and a freeze-dried microsphere type fluorescent PCR detection reagent, the detection card takes the recombinant ASFV p30 protein as a target spot, antibody detection is completed within 10 minutes, the lowest detection limit is 1: 1024, and the specificity is greater than 99%; the PCR reagent adopts a double-primer double-probe design, and the lowest detection limit is 1 copy / microliter. According to the invention, through a collaborative process of antibody screening, nucleic acid detection and result evaluation, ASFV total infection cycle detection is realized, epidemic situation discovery time is advanced to 1-3 days after infection, and early warning response time is shortened by more than 60%; the system is simple to operate and low in cost, does not need professionals and laboratories, can be deployed in basic-level scenes such as pig farms and slaughter houses, effectively solves pain points of an existing detection system, and provides key technical support for prevention and control of African swine fever.