910 results about "Classical swine fever" patented technology

The invention relates to a gene deletion attenuated African swine fever virus as a vaccine and the vaccine, and a construction method thereof. An African swine fever Chinese epidemic strain Pig/CN/HLJ/2018 is adopted, a virulence gene of the African swine fever virus is deleted by a genetic engineering technology, and the gene deletion virus of MGF360-505R deletion and joint deletion of CD2V and MGF360-505R is obtained. Experiments show that the two virus strains can provide 100% immune protection against the African swine fever Chinese epidemic virulent strains, can be used as vaccines for safe and effective prevention and control of African swine fever in China, and have great social value.

The invention discloses a four-gene-deletion weak-toxin strain for African swine fever viruses. The weak-toxin strain is the four-gene-deletion weak-toxin strain for an African swine fever virus SY18separation strain, and the following gene function protein is deleted: CD2v gene coding products and three multigene family genes( MGF360-12L, MGF360-13L and MGF360-14L ) coding products. The invention further discloses an application of the weak-toxin strain of the African swine fever viruses to preparation of vaccines for preventing or treating African swine fever. The weak-toxin strain of the African swine fever viruses can provide complete immunoprotection effect on attack of ASFV parent toxin strains, is high in safety, and is suitable for being used as vaccine candidate strains for preventing the African swine fever.

The invention relates to a composition for widely treating viral diseases. The composition contains the following raw materials, or extracts of the following raw materials, or derivatives of the extracts of the following raw materials in chemically-acceptable forms, wherein the raw materials include processed products prepared from the raw materials. The constituents of the composition are as follows: 100 parts of Baikal skullcap root, 50-150 parts of radix bupleuri and 50-150 parts of kudzu root; and according to specific circumstances, one or more selected from (but not limited to) the following raw materials can be added on this basis: radix isatidis, honeysuckle flower, Fructus Forsythiae, rhubarb, phellodendron bark, rhizoma anemarrhenae, Cyrtomium fortunei, gardenia, coptis, rhizoma cyperi, fritillaria, balloonflower and licorice root. The composition provided by the invention can be prepared into powder, liquid, paste, granules, capsules or other pharmaceutically-acceptable physical forms. The composition provided by the invention is used for preventing and treating avian influenza, epidemic hemorrhagic fever, epidemic parotitis, nameless hyperpyrexia, Newcastle diseases, swine fever, blue-ear diseases and other multiple human and animal viral diseases characterized by hemorrhage or body temperature rise, and reducing the degree of pathological damage to the body.

The invention provides CD2V protein of African swine fever (ASF) capable of being expressed with high-efficiency in a CHO (Chinese hamster ovary) cell strain. The amino acid sequence of the CD2V protein is shown in SEQ ID NO:4; a recombinant plasmid constructed by the invention is used for expressing the CD2V protein of an African swine fever virus in CHO cells; the invention further provides a recombinant CHO cell strain prepared by transfecting the CHO cells through the recombinant plasmid, the recombinant CHO cell strain can be used to prepare the CD2V protein, and the prepared protein canbe used for differential diagnosis of the African swine fever. According to the cell strain with the expression of the CD2V protein of the African swine fever, the expression quantity is high, purification is easy, the cell strain can be used for the differential diagnosis, and a solid foundation is laid for the production of subunit vaccines and diagnostic reagents of the African swine fever.

The invention relates to a vaccine production technology in the technical field of biology, in particular to a CHO cell strain which is established by utilizing a gene engineering means and is used for expressing recombinant protein PigFC-pigSCFVE2, and a preparation method and application of the recombinant protein. The recombinant fusion protein PigFC-pigSCFVE2 provided by the invention is A1) or A2) shown as follows, wherein A1) is protein of which the amino acid sequence is as shown in SEQ ID No.2, and A2) is protein which is obtained by substituting, losing and/or adding one or several amino acid residues in the amino acid sequence of the protein of the A1) and has PigFC-pigSCFVE2 activity. A monoclonal cell strain which is obtained through the method and capable of carrying out secretory expression on PigFC-pigSCFVE2 is higher in fusion protein expression quantity, fusion protein obtained through affinity separation and purification of an antibody can be combined with a monoclonal antibody, animals can be immunized, the immunity of a generated neutralizing antibody is higher than that of a present market product, the fusion protein can be used for swine classical fever preventive vaccine, and the production cost and the immunity failure loss can be reduced.

A real-time isothermal recombinase-polymerase amplification detection kit for African swine fever viruses is disclosed. A forward primer sequence for detecting the African swine fever viruses through a method provided by the kit is shown as SEQ ID NO:1, a reverse primer sequence is shown as SEQ ID NO:2 and a probe sequence is shown as SEQ ID NO:3. A real-time isothermal recombinase-polymerase amplification method provided by the invention for ASFV detection is simple and convenient in operation, rapid in reaction and low in detection cost, can be used for ASFV detection in a laboratory and on site, particularly ASFV detection in quarantine ports, airports and epidemic disease outbreak sites, and provides a novel and reliable technique support for ASF control in China.

The invention relates to a biologically safe Africa swine fever antigen multifactorial serum for ELISA (enzyme-linked immuno sorbent assay) diagnosis. A technical scheme adopted in the invention includes: adopting gene-expressed structural protein P72, K205R, P54, and A104R, conducting chemical purification, carrying out coating with a Freund's incomplete adjuvant, performing intramuscular immunization on laboratory swine in three batches, collecting swine blood after one month, separating serum, implementing serological testing, and conducting subpackaging and preservation. The serum is subpackaged into ELISA kits to undergo test according to conventional ELISA test methods.

The invention discloses a Cpf1 reagent kit for quickly detecting nucleic acid of an African swine fever virus. The Cpf1 reagent kit comprises a Cpf1 detection system suitable for quickly detecting theAfrican swine fever virus, and an immune colloidal gold test strip, wherein the Cpf1 detection system comprises specific crRNA protein, specific Cpf1 protein and a single-chain DNA(ssDNA) reporting system in accordance with a p72 gene of the African swine fever virus, the specific crRNA is one or more of crRNAs from ASFV P72 crRNA1 to ASFV P72 crRNA10, and the sequence of the specific crRNA is SEQ NO.4 to SEQ NO.13; and the single-chain DNA(ssDNA) reporting system comprises ssDNA FQreporter for fluorescence detection of a microplate reader and/or ssDNA DB reporter for detecting the immune colloidal gold test strip. According to the Cpf1 reagent kit disclosed by the invention, for the first time, the Cpf1 is used for detecting the African swine fever virus, and has the advantages of beinghigh in sensitivity, high in specificity, short in time consumption, high in flux, independent of large-scale experiment equipment and the like. The advantages enable a detection method based on the immune colloidal gold test strip developed by the invention to be conveniently used in basic laboratories and breeding enterprises to be used for performing detection, identification and diagnosis on basic quick detection of the African swine fever.

The invention relates to an ELISA kit for distinctively detecting antibodies of classical swine fever (CSF) vaccine immunity and wild virus infection and a preparation method thereof. An indirect ELISA kit or a blockage ELISA kit is formed by expressing and purifying classical swine fever virus (CSFV) non-structural protein NS3, coating a solid-phase carrier and assembling with other matched reagents. The kit has the characteristic of distinctively detecting different antibodies generated by the CSF vaccine immunity and the wild virus infection. The kit can distinctively diagnose the CSF vaccine immunity and the wild virus infection.

The invention discloses a PCR primer for detecting an African swine fever virus, a kit and application thereof, and belongs to the detection field of the African swine fever virus. According to a conserved region of an ASFV p72 gene on the GenBank, four pairs of primers are designed, the primer with strong specificity and high sensitivity is screened out from the four pairs of primers, and the primer is composed of nucleotide sequences as shown in SEQ ID No.1 and SEQ ID No.2. The invention further discloses a kit prepared from the primer and used for detecting the African swine fever virus, and a corresponding PCR detection method is established. The detection method established by the invention is more specific and sensitive in comparison with two African swine fever virus PCR detection methods recommended by OIE; and the clinical sample detection result shows that the PCR detection method disclosed by the invention is simple in operation, low in cost, good in specificity and high in sensitivity, and can be effectively applied to the screening and fast diagnosis of the African swine fever.

The invention provides an amphimorphic FQ-PCR detection reagent kit for identifying African swine fever and swine fever virus wild strains. A P72 gene of ASFV and a 5'UTR noncoding region of CSFV arerespectively used as an amplification target area, a pair of specific primers and a TaqMan MGB probe (SEQ ID NO:1-6) are designed, a real-time fluorescent quantitation PCR(FQ-PCR) technique is used, and identification and detection of ASFV and CSFV are realized. The detection reagent kit provided by the invention is suitable for detecting viral nucleic acid in samples of serum, spleen, lymph nodes, tonsil, kidney and the like of suspected ASFV or CSFV infected pigs, the sensitivity can reach 1.0*10<1>copy/[mu]L, and the detection reagent kit does not have any cross reactions with other pathogens which are liable to be in mixed infection with the ASFV and the CSFV or of which the infection symptoms are similar such as PRRSV, PRV, PCV2, PPV, JEV and HPS.

The invention discloses a hybridoma cell line of a monoclonal antibody against African swine fever virus and the secreted monoclonal antibody thereof. The preparation method of the invention comprises the following steps: preparing a recombined P30 soluble antigen by prokaryotic expression; immunizing a BALB/c mouse; and finally fusing, screening and cloning by a hybridoma technology to obtain the hybridoma cell line which can stably secrete the monoclonal antibody against African swine fever virus P30 protein. The invention further discloses a method for preparing the monoclonal antibody with the cell line, an antibody purification method and a labeling method for horseradish peroxidase of the antibody. The monoclonal antibody can be used in detecting the African swine fever viral antibody in pig serum.

The present invention relates to African swine fever virus (ASFV) peptides and/or polypeptides as well as immunogenic fragments thereof, corresponding encoding AFSV oligonucleotides and/or polynucleotides as well as immunogenic fragments thereof, immunogenic compositions, vaccines and uses thereof.

The invention provides a CSFV antibody detection system and a preparation method thereof. A coating antigen of the detection system contains CSFV E2 protein and Erns protein. The CSFV E2 protein and Erns protein are recombinant proteins expressed by eucaryon, correct spatial conformation and posttranslational modification process can be guaranteed, antigen is capable of effectively combining with the antibody in serum, and specificity, sensitivity and repeatability of detection can be increased. The system and the method can be sued for diagnosis of CSFV antibody in prevention and control of CSFV as well as immunization evaluation of a CSFV vaccine.

The invention discloses a traditional Chinese medicine composition for treating a disease of swine fever, which relates to the technical field of veterinary medicines in biomedicine. The traditional Chinese medicine composition for treating the disease of the swine fever is prepared from the following raw materials according to parts by weight: 12 to 16 of rhizoma coptidis, 18 to 20 of baikal skullcap root, 15 to 18 of yellow-corktree bark, 18 to 20 of rhubarb, 15 to 18 of honeysuckle, 15 to 17 of gardenia, 12 to 14 of gypsum, 15 to 18 of rhizoma anemarrhenae, 20 to 30 of dandelion, 14 to 18 of garden burnet, 22 to 28 of white peony root, 15 to 18 of tuckahoe, 22 to 28 of officinal magnolia, 12 to 15 of poncirus trifoliata, 13 to 15 of dark plum fruit, 18 to 20 of realgar and 22 to 28 of licorice. The traditional Chinese medicine composition for treating the disease of the swine fever has the effects of clearing heat, expelling toxin, removing heat from the blood, purging intense heat, removing dampness by diuresis, relieving restlessness, washing bowels, removing the stagnancy of the fu-organs, regulating qi, strengthening the body resistance, regulating the functions of the liver and the spleen and the like. The traditional Chinese medicine composition for treating the disease of the swine fever has the characteristics that the traditional Chinese medicine composition for treating the disease of the swine fever is suitable for treating the disease of the swine fever, the curative effect is high and the like. The traditional Chinese medicine composition for treating the disease of the swine fever not only can be used for treating the disease of the swine fever with various epidemic situations but also can be used for preventing the disease of the swine fever.

The invention discloses a traditional Chinese medicine for treating swine fever, which is characterized in that the traditional Chinese medicine is prepared by the following components according to weight portions: 30 to 50 portions of realgar, 20 to 50 portions of fineleaf schizonepeta herb, 20 to 50 portions of honeysuckle flower, 20 to 50 portions of radix bupleuri, 20 to 50 portions of radix puerariae, 20 to 50 portions of baical skullcap root, 20 to 50 portions of akebia stem, 20 to 50 portions of radix isatidis, 20 to 50 portions of liquoric root, 20 to 50 portions of indigowoad leaf and 20 to 50 portions of dried ginger. The traditional Chinese medicine has more obvious effect for preventing and treating swine fever.

The invention belongs to the field of sanitary examination, and relates to a LAMP kit for testing classical swine fever virus and an establishing method and an application thereof. The kit contains a test system which is composed of the LAMP reaction liquid of six LAMP primers. The tests prove that the kit of the invention has good specificity and sensitivity, fast amplification speed, high efficiency and simple and convenient identification. The test system of the invention can rapidly and conveniently test the classical swine fever virus in high-efficiency, high-specificity and high-sensitivity under the isothermal condition of 65 DEG C without complicate instruments, can better satisfy the spot tests for the classical swine fever virus, provides a novel technical platform for food safety testing, can better meet the urgent requirements for the spot testing of foot and mouth diseases at present, is used for the spot testing of import and export quarantine, food sanitary departments, animal breeding farms, etc, and is easy to be popularized in a wide range.

The invention discloses an RPA (recombinase polymerase amplification) primer and a detection kit for rapidly detecting African swine fever viruses. Target genes can be effectively amplified, specificity is 100%, detection sensitivity is 102 copy/reaction, and sensitivity is equivalent to that of fluorescent quantitative PCR (polymerase chain reaction). Cross reaction between the RPA amplificationprimer and one of classical swine fever viruses, vesicular exanthema swine viruses I, porcine reproductive and respiratory syndrome viruses, porcine circoviruses and the like is omitted. A RPA isothermal amplification system is rapidness in reaction and wide in temperature range, effective amplification of the target genes can be achieved at the temperature of 38-46 DEG C, and the detection kit can rapidly, efficiently and sensitively detect the African swine fever viruses and has the advantages that the kit is simple to operate, high in specificity, safe and free from pollution, reaction results are easily observed and the like. Effective technical means are provided for on-site rapidness detecting and screening of infection nucleic acid of the African swine fever viruses, and the RPA amplification primer has great significance for control of infection spreading of the African swine fever viruses in China and inspection and quarantine in infected areas and entry and exit ports.

The invention relates to a fluorescence quantitative PCR rapid diagnosis kit for specifically detecting the Hog cholera virus and wild virus infection and an application method thereof, belonging to the virus nucleic acid detection field. The fluorescence quantitative PCR rapid diagnosis kit is applied to the rapid quantitative detection of the CSFV wild virus clinically and in scientific research and can remove the false positive caused by the CSFV HCLV vaccine immunization. Compared with the prior art, the fluorescence quantitative PCR rapid diagnosis kit has the advantages that: the Hog cholera virus and wild virus infection and the vaccine immunization can be verified and diagnosed, the problem of the incapability of removing the false positive caused by the vaccine immunization is solved; the specificity is good, the high specificity hybridization double control of the primer high specificity amplification and the fluorescence probe is realized, the accuracy is high, the false positive is low; the sensitivity is high; the detection speed is high, only one hour is needed and two to three hours are needed when the extraction process of the nucleic acid and the preparation process of the cDNA are added; and the post treatment is not necessary, the processes such as hybridization, electrophoresis and photo are not necessary and no pollution is caused.

The invention provides a method for establishing a recombinant adenovirus vector with Africa swine fever EP153R and P54 gene coexpression and packaging adenovirus and belongs to the technical field ofgene engineering. According to the adenovirus vector of coexpression of the EP153R gene and the P54 gene, with a pShuttle-CMV eukaryotic expression vector as a basis, the CTLA4, EP153R and P54 genesare introduced at multiple cloning sites. The invention further provides recombinant adenovirus with coexpression of the EP153R and P54 genes. The established pAD-Shuttle-CMV-CTLA4-EP153R-P54 vector is used for achieving the adenovirus packaging process, the adenovirus which can directly infect animals or eukaryocyte is obtained, the aim of coexpression of the EP153R and P54 genes in eukaryocyte is achieved, and the basis is laid for research on adenovirus vaccines of coexpression of the EP153R and P54 genes.