171 results about "Classical swine fever" patented technology

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 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 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 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.

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 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 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 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.

The invention discloses a quadruple real-time fluorescent quantitative PCR (Polymerase Chain Reaction) primer probe combination for detecting African swine fever virus and application. The primer probe combination comprises a first primer probe combination, a second primer probe combination, a third primer probe combination and a fourth first primer probe combination, the primer probe is capable of specifically targeting ASFV p72 gene, CD2v-I gene, CD2v-II gene and I177L gene, and the detection kit developed based on the primer probe combination can rapidly diagnose African swine fever virus and identify CD2v gene I and II type and CD2v and I177L gene deleted strains. The kit has extremely high sensitivity, the lowest detection limit can reach 8 copies / mu L, it is ensured that trace virus DNA can be detected, and missing detection is avoided.

The invention belongs to the technical field of biology, and particularly relates to a genotyping dot matrix chip for detecting African swine fever viruses as well as a construction method and application of the genotyping dot matrix chip. Firstly, the invention provides a single nucleotide polymorphism site combination for accurately detecting African swine fever gene type I, type II and I / II recombinant viruses; secondly, the invention provides a genotyping dot matrix chip for detecting the African swine fever virus, and the chip can simultaneously realize accurate typing, epidemic and variation monitoring and I / II recombinant virus recombinant gene identification of a high-throughput detection sample.

This invention relates to the field of biotechnology, and aims to provide a monoclonal antibody 7A12 for identifying genotype 2 classical swine fever, along with its preparation and application methods. This monoclonal antibody contains an Ig domain V of the antibody heavy chain. H CDR1, V H CDR2 and V H CDR3, and the Ig domain V of the antibody light chain L CDR1, V L CDR2 and V L CDR3; wherein, the V H CDR1, V H CDR2 and V H The amino acid sequences of CDR3 are shown in SEQ ID NO:1-3, respectively; the V L CDR1, V L CDR2 and V L The amino acid sequences of CDR3 are shown in SEQ ID NO:4-6. The antibody provided by this invention has advantages such as high titer, strong specificity, and high affinity; it also has high sensitivity and specificity, and shows no cross-reactivity with type 1 classical swine fever vaccine strains. Serologically, it can differentiate and diagnose infection and vaccine immunization with genotype 2 classical swine fever wild-type strains. Furthermore, it can be used to develop a blocking ELISA kit for identifying genotype 2 classical swine fever, effectively diagnosing and purifying genotype 2 classical swine fever virus.

The invention relates to crRNA for detecting an African swine fever virus gene type I or type II and a kit. The kit comprises crRNA-1 for detecting the African swine fever virus gene type I and crRNA-2 for detecting the African swine fever virus gene type II. And the like. The kit has high specificity on African swine fever virus gene type I and gene type II, has no cross reaction with porcine reproductive and respiratory syndrome virus, porcine parvovirus, porcine circovirus and porcine Delta coronavirus, has high detection sensitivity, and can detect 4 copies / [mu] L of ASFV gene type I plasmid and 2 copies / [mu] L of ASFV gene type II plasmid. The kit provided by the invention is simple to operate, does not need large-scale instruments and equipment, and can be used for detecting the African swine fever virus in a field environment; the whole detection process is completed within 40 minutes.

The invention discloses a pig IFNAR2 protein mutant, which is characterized in that the 399th site in the amino acid sequence of the pig IFNAR2 protein is mutated from glycine to alanine, and the amino acid sequence of the pig IFNAR2 protein is as shown in SEQ ID NO. 1. The invention provides a specific cleavage site of ASFV protease pS273R, and the site is mutated through a gene editing technology so as to block the cleavage of pig IFNAR2 by the pS273R and improve the ability of pigs to resist African swine fever virus infection. By blocking ASFV from damaging type I IFN mediated JAK-STAT signal activation, on the basis of retaining pig autoimmune performance, higher specificity and higher efficiency are achieved, negative effects of genetic engineering on pig growth performance and reproductive performance can be greatly reduced, and biosafety is higher.

The application belongs to the technical field of biology and particularly relates to a nano antibody against African swine fever virus K205R protein and application. The application provides a nano antibody KR-6 against African swine fever virus K205R protein, wherein the nano antibody KR-6 is obtained through construction and screening of a phage display nano antibody library, is a heavy chain antibody variable region fragment, and has an amino acid sequence shown in SEQ ID NO. 1. The nano antibody has high affinity to African swine fever virus K205R protein and African swine fever virus, can specifically recognize African swine fever virus, and can be used for diagnosis, prevention or treatment of African swine fever.

This utility model relates to the field of African swine fever detection technology and discloses an African swine fever detection device, including a detection platform and a detection slot opened at the top of the detection platform. The mixing component includes a mixing platform and a mixing hole slot opened at the top of the mixing platform. A dilution bottle is installed inside the mixing hole slot, and an internal threaded cap is installed at the top of the dilution bottle. A mixing cap is installed at the top of the mixing platform and at the top of the internal threaded cap. The capacity of the mixing component can be adjusted according to different detection items. By rotating the lower tube, the inner tube moves down from inside the outer threaded tube, and the capacity inside the upper tube and the lower tube increases. The enlarged dilution bottle is inserted into the mixing hole slot. The outer threaded tube is rotated downward, and the space for placing the dilution bottle increases. After increasing the space capacity for mixing blood samples and diluents, the space for placing dilution bottles for mixing is also deepened, which facilitates the mixing of different blood samples and diluents.

The invention belongs to the technical field of genetically engineered bacteria, and particularly discloses an African swine fever oral vaccine saccharomyces cerevisiae engineered bacterium as well as a construction method and application thereof. A free plasmid system is adopted to transform saccharomyces cerevisiae, and an engineering strain for displaying ASFV p30 / p54 antigens is constructed. The matched plasmid system has the advantages of being simple and convenient to operate and short in construction period, construction and transformation of recombinant plasmids can be completed in a short time, and the research and development efficiency of vaccine candidate strains is remarkably improved. Meanwhile, the free plasmid can be stably inherited and copied in a yeast host, so that the expression stability of a target antigen in a passage process is guaranteed, and large-scale fermentation preparation is facilitated. After oral immunization, the saccharomyces cerevisiae engineering bacterium constructs a system-mucous membrane dual defense barrier in a tested animal pig body, not only induces high-level system IgG, but also induces remarkable sIgA mucous membrane immunity in intestinal tracts, and provides a new idea and technical reserve for research and development of ASFV subunit vaccines.

The present invention discloses a competitive chemiluminescence detection kit for detecting African swine fever virus (ASFV) p30 antibodies and its application. The present invention utilizes the principle that LgBiT and HiBiT complement each other to form a luciferase-catalyzed substrate furimazine to produce chemiluminescence. After ASFVp30 monoclonal antibody 16-5E7E8 is fused with HiBiT for expression, ASFVp30 antibodies in serum are detected using competitive chemiluminescence. The kit of the present invention is not only highly sensitive and specific for detecting ASFVp30 antibodies, but also simple to operate and has a short reaction time. It only takes 20 minutes to quickly and effectively detect whether the serum to be tested contains ASFVp30 antibodies. It has a good application prospect in ASFV antibody detection and provides technical means for the comprehensive prevention and treatment of ASF.

The application belongs to the technical field of animal vaccines, and particularly relates to an O-A-Dex-Ap for African swine fever immunization. Based on the good biocompatibility of Dextran, the application uses the Dextran as a nanoparticle to load a coupled CTL epitope peptide. Preliminary experimental results show that, compared with the epitope peptide alone, the O-A-Dex-Ap after loading and coupling can more effectively promote antigen presentation and activate more persistent CTL immune response, and has good application potential. Based on the results, a good technical foundation can be laid for subsequent preparation of African swine fever and other disease vaccines.

The present application belongs to the field of molecular biology detection, and particularly relates to a composition, method and use for simultaneous detection and differentiation of swine fever virus, psittacosis chlamydia, porcine reproductive and respiratory syndrome virus and porcine circovirus type II. The composition for simultaneous detection provided by the present application can realize rapid detection of four pathogen target genes in one tube of reaction solution by using multiplex fluorescence RPA detection technology, and has the advantages of high throughput, short time and low cost. On one hand, the present application can accurately detect pathogens and analyze mixed infection of pathogens, so as to achieve the effect of precise medication, reduce the use of antibiotics, save costs, and reduce the cost and increase the benefit of enterprises, and on the other hand, compared with fluorescence PCR, the detection time is greatly shortened, and the purpose of rapid detection is achieved.

The application discloses a single-domain antibody against African swine fever virus E165R protein and a screening method and application thereof, relates to the technical field of bioengineering, and provides the single-domain antibody against African swine fever virus E165R protein and a specific amino acid sequence, a gene coding the single-domain antibody, an expression vector comprising the gene and a bacteriophage expressing the gene, and applies the single-domain antibody against African swine fever virus E165R protein to immunological detection of African swine fever virus. The single-domain antibody provided by the application has strong specificity, high sensitivity and strong neutralization activity, and can be used in daily immunological detection of epidemic diseases and future system construction for prevention and control of epidemic diseases.

This invention belongs to the field of bioengineering technology, specifically relating to the construction of a double-gene-deleted attenuated African swine fever virus strain and its application as a vaccine. This invention combines the deletion of the MGF505-7R and I267L genes in the ASFV CN / GS 2018 virus, reducing the virulence of the parent strain and obtaining an attenuated African swine fever vaccine strain. The attenuated African swine fever vaccine strain completely attenuates pigs after immunization, with 100% survival rate, and provides immune protection against challenge with the virulent ASFV CN / GS / 2018 strain. It can serve as a safe and effective candidate vaccine for the prevention and control of African swine fever, possessing significant social value.

The application discloses an African swine fever H240R epitope peptide, a monoclonal antibody and application thereof, and relates to the technical field of molecular immunology. The amino acid sequence of the African swine fever H240R epitope peptide is shown as SEQ ID NO. 19 or SEQ ID NO. 20. The monoclonal antibody is monoclonal antibody 11C1A6 or monoclonal antibody 38A3B2; the amino acid sequences of the heavy chain variable region and the light chain variable region of the monoclonal antibody 11C1A6 are shown as SEQ ID NO. 9-10, respectively; the amino acid sequences of the heavy chain variable region and the light chain variable region of the monoclonal antibody 38A3B2 are shown as SEQ ID NO. 11-12, respectively. The African swine fever H240R epitope peptide is identified, and the monoclonal antibody which is specifically combined with the African swine fever H240R protein is prepared, thereby laying a foundation for preparing an African swine fever polypeptide vaccine and a detection kit.