30 results about "Aphthovirus" patented technology

The invention discloses a sheep aphthovirus Asial type multi-epitope recombinant vaccine and a preparation method thereof. The recombinant vaccine comprises the following ingredients: a protein encoded by aphthovirus multi-epitope genes and carrier protein fusion genes, and an aphthovirus 3D protein. The preparation method comprises the following steps: respectively diluting the protein encoded by the aphthovirus multi-epitope genes and the carrier protein fusion genes and the aphthovirus 3D protein; then, uniformly mixing the diluted proteins; adding auxiliary agents into the mixture; carrying out emulsification; and obtaining the sheep aphthovirus Asia1 type multi-epitope recombinant vaccine. Animal model and animal immune efficiency experiments show that the sheep Asial epitope recombinant vaccine of the invention can generate overall immunoprotection reaction, injected immune sheep can be induced to generate high-level neutralizing antibodies, and can also induce the cellullar immunologic response. The recombinant vaccine of the invention can effectively protect animals from the virulent strain attack of the aphthovirus.

The invention discloses an RT-RPA detection reagent kit for quickly detecting pest des petits ruminant viruses and application thereof. The reagent kit comprises a pair of primers and a probe. The sequences of the primers are as shown in SEQ ID NO.1 and SEQ ID NO.2, and the sequence of the probe is as shown in SEQ ID NO.3. Experiments prove that the reagent kit can specifically detect the pest des petits ruminant viruses and does not amplify sheep pox viruses, goat pox viruses, contagious pustular dermatitis sheep viruses, foot and mouth disease viruses and other important sheep infection viruses under the same condition, and it is illustrated that the method has good specificity. Sensitivity experiments show that the RT-RPA primers and the probe for quickly detecting the pest des petits ruminant viruses can at least detect 150 copied templates at the temperature of 40 DEG C and under the condition that amplification is conducted for 20 min and have high conformity with RT-qPCR. Accordingly, the reagent kit can quickly, efficiently and sensitively detect the pest des petits ruminant viruses, and an effective technical means is provided for identification and diagnosis of the pest des petits ruminant viruses.

The invention relates to a method for constructing porcine fmd recombinant immune complex peptide and an application thereof, which belongs to the technical field of biological engineering. The immune multi-peptide adjuvant is fused with aphthovirus novel auxiliary T cell epiposition (I-S-I-S-E-I-K-G-V-I-V-H-K-I-E-T-I-L-F) and Bursin mimic peptide (T-P-N-L-K-H-G) by a flexible Linker. A fusion gene is inserted into an expression carrier to be converted into colon bacillus, gene engineering strains which express the porcine fmd recombinant immune complex peptide with high efficiency are obtained, and the fusion protein of the porcine fmd recombinant immune complex peptide is obtained after liquid culture and purification; the thioredoxin in the fusion protein is removed by enterokinase with His label on the N end, and the single porcine fmd recombinant immune complex peptide can be obtained after affinity chromatography purification with very high sample purity. The recombinant immune complex peptide can serve as the novel immune multi-peptide adjuvant of the foot-and-mouth disease vaccine. Animal immune experiments prove that the method has high safety but no toxicity and side effect, and can effectively enhance the immunity of organisms and cells and the immunity level of body fluid.

The invention discloses a real-time fluorescent quantitative PCR (polymerase chain reaction) detecting reagent kit for an orf virus and belongs to the technical field of viral molecular biology detection. The real-time fluorescent quantitative PCR detecting reagent kit for the orf virus comprises a pair of primers of nucleotide sequences shown in SEQ ID NO:1-2 and a TaqMan probe of a nucleotide sequence shown in SEQ ID NO:3. The detecting reagent kit can quickly and accurately detect the orf virus from samples and is high in detecting sensitivity and simple and convenient to sample, detecting efficiency is greatly improved, pollution can be effectively prevented, and the detecting reagent kit is of practical significance in the field of inspection and quarantine of import and export animals.

Equine rhinovirus 1 (ERhV1) is a respiratory pathogen of horses which has an uncertain taxonomic status. The nucleotide sequence of the ERhV1 genome and amino acid sequence have been substantially determined (FIG. 2). The predicted polyprotein was encoded by 6,741 nucleotides and possessed a typical picornavirus proteolytic cleavage pattern, including a leader polypeptide. The genomic structure and predicted amino acid sequence of ERhV1 were more similar to those of foot-and-mouth disease viruses (FMDV), the only members of the aphthovirus genus, than other picornaviruses. Nucleotide sequences coding for the complete polyprotein, the polymerase, and VP1 were analyzed separately. The phylogenetic trees confirmed that ERhV1 was more closely related to aphthoviruses than to other picornaviruses. Virion proteins and virus-like particles are described and probes, primers, antigens, vectors, diagnostics and tests developed.

The invention discloses a sheep pox and aphtha bigeminy cell attenuated vaccine and a preparation method and application thereof. The effective ingredient of the sheep pox and aphtha bigeminy cell attenuated vaccine is sheep pox virus and contagious pustular dermatitis virus cell passage attenuate virus or passage virus thereof which are separated through the inventor and obtained after passage attenuate culture, wherein the sheep pox virus cell passage attenuate virus is Sheeppox Virus GS-WW 2010 F44 strain, the preservation number of CCTCC NO: V201504, the sheep pox virus (contagious pustular dermatitis virus) cell passage attenuate virus is Orf Virus HB-TS09F65 strain, and the preservation number is CCTCC NO:V201406. Toxin attacking protection test results show that the sheep pox and aphtha bigeminy cell attenuated vaccine is safe and capable of producing effective immune protection force. Accordingly, the sheep pox and aphtha bigeminy cell attenuated vaccine is of an important practical significance in researching safe and efficient attenuated vaccine and preventing and controlling sheep pox and aphtha bigeminy.

The invention is directed to an adenoviral vector comprising at least one nucleic acid sequence encoding an aphthovirus antigen and/or a cytokine operably linked to a promoter. The adenoviral vector is replication-deficient and requires at most complementation of both the E1 region and the E4 region of the adenoviral genome for propagation. The invention also is directed to a method of inducing an immune response in a mammal comprising administering to the mammal a composition comprising the aforementioned adenoviral vector.

The invention relates to a method for rapidly separating a sore mouth disease virus and in particular relates to a method for separating the sore mouth disease virus from peripheral blood of ill sheep and recessive virus-carried sheep by utilizing bovine testicle cells. The sore mouth disease virus is separated from the peripheral blood of ill sheep suffering from the sore mouth disease and the recessive virus-carried sheep by utilizing the bovine testicle cells, a convenient and rapid method is provided for separating the sore mouth disease, and convenience is brought to production and scientific research. The method for rapidly separating sore mouth disease virus disclosed by the invention comprises the following steps: (1) collecting and detecting a sore mouth disease virus sample; (2) treating the sample; and (3) separating the sore mouth disease virus.

The invention provides an orf virus attenuated strain and an application thereof. The attenuated strain is an orf virus attenuated strain QFnm2015, the preservation institution is the China Center forType Culture Collection, and the preservation number is CCTCC NO: V201903. According to the invention, the isolated orf virus is proliferated and cultured on lamb testicular cells, and the orf virusattenuated strain is cultured through continuous passage. The isolated strain of the orf virus provides a high-quality material basis and selection space for developing an effective vaccine of endemicORFV, and has important significance for prevention and treatment of orf.

The invention provides a recombinant DNA vaccine and a recombinant plasmid thereof, the DNA vaccine recombinant plasmid comprises an antigen coding sequence and a plasmid vector, the antigen coding sequence comprises an orf antigen coding sequence and a sheep pox antigen coding sequence which are connected in series, and an orf antigen and a sheep pox antigen are expressed as independent antigen proteins. The invention provides construction of a double-gene recombinant DNA (Deoxyribose Nucleic Acid) vaccine of an orf virus gene and a sheep pox virus gene and analysis of a mouse immune effect. According to the invention, an orf virus B2L (011) gene and a sheep pox virus P32 gene are selected, the two genes are connected in series by using a self-cleaving peptide P2A, and the self-cleaving peptide P2A generates self-cleaving in eukaryotic cells, so that two antigen proteins are independently expressed on an eukaryotic expression vector pcDNA3.1 (+). The recombinant DNA vaccine prepared by the invention also has a better immune effect after being used for immunizing a mouse, and can be used as a reference of a candidate vaccine.

The invention discloses a kit of double RPA detection of aphthovirus and vesicular stomatitis virus. The kit includes an aphthovirus RPA detection primer set consisting of nucleotide sequences shown as SEQ. ID. NO. 1 and SEQ. ID. NO. 6 and a vesicular stomatitis virus RPA detection primer set consisting of nucleotide sequences shown as SEQ. ID. NO. 8 and SEQ. ID. NO. 11. Based on a recombinase polymerase amplification method, the kit takes the cDNA formed by RNA reverse transcription as a template and performs amplification reaction under optimized reaction temperature and time, and detection results can be obtained through nucleic acid gel electrophoresis. The kit can be used to simultaneous detect aphthovirus and vesicular stomatitis virus, is simple in operation and has high sensibility and specificity, therefore rapid detection kit can be provided for on-site screening of pathogens.

The invention relates to a virus isolation method for a low-content sore mouth virus sample. The traditional sore mouth virus is separated from incrustation and oral lip scurf of an ill goat, the isolation period is long, lesion is caused after 5 generations of blind passage, the pathological sample source range is narrow, and the research work of the sore mouth virus is limited. The method disclosed by the invention comprises the following steps: performing centrifugal filtration treatment on samples such as saliva, breast milk and internal organs by using a special Virus holder solution, inoculating the cattle primary testis cells with the samples for culturing, and discarding the venom; adding cell maintenance medium for culturing, and collecting the virus; and repeatedly freezing-thawing and collecting the cell culture, inoculating the cattle testis cells from the third generation within 72 hours, and observing the cytopathic effect. The samples are treated by using the special Virus holder solution, the sore mouth virus can be isolated from a sample with extremely low virus content, the material taking range is wide, the virus isolation period is short, the stability is high, and the integrity and activity of the virus can be well maintained.

The invention discloses an environment-friendly aphthovirus purifying compound percarbamide effervescent granule disinfectant. The disinfectant is prepared from the following ingredients in percentage by weight: 1 to 5 percent of percarbamide, 10 to 20 percent of potassium hydrogen persulfate, 5 to 10 percent of citric acid, 2 to 5 percent of fumaric acid, 10 to 29 percent of sodium bicarbonate, 5 to 10 percent of lauryl sodium sulfate, 0.1 to 0.9 percent of hydrochloric acid, 0.1 to 1.9 percent of sulfuric acid, 1 to 5 percent of phosphoric acid, 2 to 5 percent of lactic acid, 5 to 10 percent of vitamin C, 10 to 27 percent of maltodextrin and 5 to 15 parts of sodium chloride. The prepared disinfectant has the advantages that the environment is protected; the use cost is low; the dosage is novel. The contained raw materials of the disinfectant can be degraded into carbon, hydrogen and oxygen under the natural conditions after the use; soil pollution and water pollution cannot be generated. The disinfectant provided by the invention has low cost, and has the exact killing curative effect on the aphthovirus.

The invention discloses a high-throughput micro-fluidic LAMP chip for detecting various original bacteria of goat epidemic diseases and a detection method, and belongs to the technical field of biological chips. The LAMP chip comprises an LAMP amplification reagent, a DNA template, a quality control primer positive control PC, a quality control primer negative control NC and a chip probe of goat plague pathogenic bacteria; a detection chip is arranged in an isothermal amplification micro-fluidic LAMP chip analyzer; and the chip probe comprises nucleotide sequences as shown in SEQ ID NO1-27, and is used for detecting specific nucleic acid fragments of brucella melitensis, peste des petits ruminants virus, mycoplasma ovipneumoniae, goatpox viruses and orf virus. According to the invention, detection can be completed within 20-50 minutes; furthermore, the detection limit is 1.5*10<-2> ng/mu L; main pathogenic bacteria of goats can be monitored in real time; the method has the characteristics of simplicity in operation, rapidness in detection, high sensitivity and high specificity; and convenience is brought to detection of the main pathogenic bacteria of the goats.

The invention discloses a sheep fetal skin fibroblast and a separation method and application thereof. The preservation number of the fibroblast is CCTCC No: C2019202. In the separation process of the sheep fetal skin fibroblast SFSFS, digestive juice containing 0.05% trypsin and 0.02% EDTA and IV-type collagenase are adopted for treating sheep fetal skin tissue, and then digestive juice containing 0.01-0.05% trypsin-EDTA is used for treating cells; and 3% fetal calf serum and 10-40ng/ml h-EGF are adopted for replacing 10% fetal calf serum to culture the sheep fetal skin fibroblast, the use amount of the fetal calf serum is greatly reduced, the SFSFS culture cost and the aphtha virus production cost are reduced, and resources are saved. The cell strain can obviously increase the separation rate of aphtha viruses and shorten the separation time, the separated aphtha virus titer and virus copy number are higher, and a tool can be provided for large-scale reproduction of the aphtha viruses and virus infection pathogenesis research.