30 results about "Aphthovirus" patented technology

A determination of the nucleotide sequence for the genome of Equine rhinovirus 1 (ERhV1), a horse-respiratory pathogen of heretofor uncertain taxonomic status, reveals a structure and predicted amino acid sequence that are more similar to those of foot-and-mouth disease viruses, the only members of the aphthovirus genus, than of other picornaviruses. These insights inform an understanding of ERhV1 virion proteins and virus-like particles, as well as the design of probes, primers, antigens, vectors, diagnostics and tests of relevance to ERhV1.

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.

A multiplex-PCR (polymerase chain reaction) kit for detecting six viruses of sheep and goats simultaneously comprises six pairs of specific primers, wherein the primers used for detection of the bluetongue virus are BTV-F and BTV-R respectively; the primers for detection of the foot and mouth disease virus are FMDV-F and FMDV-R respectively; the primers for detection of the peste des petits ruminants virus are PPRV-F and PPRV-R respectively; the primers for detection of the sheep pox virus are SPPV-F and SPPV-R respectively; the primers for detection of the goat pox virus are GTPV-F and GTPV-R respectively; the primers for detection of the sore mouth disease virus are ORFV-F and ORFV-R respectively. The sequences of the primers are nucleotide sequences shown in SEQ ID NO: 1-12. The multiplex-PCR kit can be used for detecting nucleic acid of the six viruses including the bluetongue virus, the foot and mouth disease virus, the peste des petits ruminants virus, the sheep pox virus, the goat pox virus and the sore mouth disease virus in sheep and goat clinical samples and has the characteristics of high sensitivity, high specificity, simple operation and reduction of detection time.

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 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 relates to a preparation method of a permanent cell line for multiplying an orf virus. The aim that the virus can be stably multiplied in a large amount is achieved by inoculating a calf testicular cell line to the orf virus and a stable cell environment and a standardized culture system for developing and producing an attenuated vaccine of the orf virus are provided. The preparation method comprises the following steps of: firstly, acquiring, separating and culturing a calf testicular cell; secondly, permanently establishing the calf testicular cell; and thirdly, carrying out multiplication culture on the orf virus in the cell line.

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 relates to a permanent cell line for ecthyreosis virus multiplication. The name of the cell line is a goat testis cell GT26, the cell line is preserved at the China Center for Type Culture Collection on March 1, 2017, and the preservation number is CCTCC NO:C201734. An establishment method of the permanent cell line comprises the steps of (1) permanent establishment of the goat testis cell; (2) low-serum domestication of the goat testis cell; and (3) cultivation of an ecthyreosis virus in a newborn goat testis cell line. The ecthyreosis virus is grafted by using the goat testis cell line, thereby achieving the target of stable virus multiplication and providing a stable cell environment and a standardized culture method for development and production of an attenuated vaccine and an inactivated vaccine of the ecthyreosis virus.

The invention provides a method for preparing a yeast cDNA library of sheep testis fibroblast membrane system, which comprises preparing sheep testis fibroblasts; extracting RNA from sheep testis fibroblasts; synthesizing cDNA and homogenizing the cDNA library; cDNA and pPR3-N carrier Ligation, transformation of the ligation product into host cell DH5α, obtaining the yeast cDNA library of the sheep testis fibroblast membrane system, and performing cloning verification; screening the yeast cDNA library of the sheep testis fibroblast membrane system. The cDNA library capacity of the present invention is 7.5 × 10 5 , the size of the fragments is 200‑2000bp, the average length is 1000bp, and the recombination rate of the library is 100%. The library was screened by using the membrane protein of aphthus virus, which proved that the library was a high-quality library.

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 a real-time fluorescent quantitative PCR detection kit for aphthous ulcer virus, which belongs to the technical field of virus molecular biology detection. The real-time fluorescent quantitative PCR detection kit for aphthus virus of the present invention comprises a pair of primers with the nucleotide sequence shown in SEQ ID NO: 1-2 and a TaqMan probe with the nucleotide sequence shown in SEQ ID NO: 3. The detection kit can quickly and accurately detect the aphthus virus in the sample, and has high detection sensitivity, simple sampling, greatly improved detection efficiency, and can effectively prevent pollution. It has great practical significance in the field of import and export animal inspection and quarantine.

The invention discloses a goatpox virus-orf virus combined cell attenuated vaccine and its preparation method and use. The goatpox virus-orf virus combined cell attenuated vaccine contains an active component which comprises goatpox and contagious pustular dermatitis virus cell passage-attenuated viruses or passaged viruses separated by the inventor and subjected to passage attenuation culture, wherein the goatpox virus cell passage-attenuated virus is a Goatpox Virus HN-XY2010F43 strain and has a preservation number of CCTCC NO: V201503 and the contagious pustular dermatitis virus cell passage-attenuated virus is an Orf Virus HB-TS09F65 strain and has a preservation number of CCTCC NO: V201406. A toxin counteracting protection test result shows that the goatpox virus-orf virus combined cell attenuated vaccine is safe and can produce an effective immunization protection function. The goatpox virus-orf virus combined cell attenuated vaccine and its preparation method have important practical meanings for development of safe and efficient attenuated vaccines and large-scale production of the goatpox virus-orf virus combined cell attenuated vaccine for preventing and controlling goatpox and orf from the disclosed attenuated strain.