43 results about "Bluetongue disease" patented technology

The invention discloses a duplex fluorescent RT-LAMP detection group and kit for visually identifying foot-and-mouth disease virus and Bluetongue virus and an application of the detection group and the kit. The duplex fluorescent RT-LAMP detection group comprises two groups of specific primers and probes, the first group of specific primers and probes comprise FMDV-F3, FMDV-B3, FMDV-FIB(F1c-F2), FMDV-BIP(B1c-B2) and FMDV-probes, the second group of specific primers and probes comprise BTV-F3, BTV-B3, BTV-FIB(F1c-F2), BTV-BIP(B1c-B2) and BTV-probes, and the two groups of specific primers and probes are as shown in a sequence table SEQ ID No.1 to SEQ ID No.10. The duplex fluorescent RT-LAMP detection group can identify and diagnose the foot-and-mouth disease virus and the Bluetongue virus inthe same reaction tube and has the advantages of good specificity, high sensibility, less pollution, convenience, rapidness and the like, a detection result can be directly observed by naked eyes andis judged according to colors of reaction products, and the detection group can be used for basic-level quarantine with poor conditions.

The invention aims at providing a primer and a probe for detecting peste des petits ruminants virus by virtue of a RPA (Recombinase Polymerase Amplification) technique. According to the primer and the probe, a forward primer sequence is represented by SEQ ID NO:1, a reverse primer sequence is represented by SEQ ID NO:2, and a probe sequence is represented by SEQ ID NO:3. According to a method, a large number of primers and probes are designed according to genomic sequences of the peste des petits ruminants virus, and a pair of primer and probe combinations capable of rapidly detecting nucleic acid of the peste des petits ruminants virus are screened out; obvious amplification curves can be obtained by carrying out rapid detection by virtue of the primer and the probe and taking nucleic acid RNA of the peste des petits ruminants virus of domestically separated Tibet strains and Xinjiang strains as a template, and no amplification curve is obtained through a reaction by taking other pathogenic nucleic acids such as foot and mouth disease virus, contagious pustular dermatitis virus, mycoplasma capricolum and bluetongue disease virus as the template.

The invention discloses an anti-sheep virus disease yolk antibody feed additive, an injection and a preparation method thereof. The anti-sheep virus disease yolk antibody feed additive and the injection respectively are extracted and prepared by the collected yolk antibody of the immunized laying hens which are immunized by the single vaccine which is prepared form sheep pustule virus, sheep bluetongue disease virus and sheep foot and mouth disease virus or the combined vaccine which is prepared from sheep pustule virus and / or sheep bluetongue disease virus and / or sheep foot and mouth disease virus. The anti-sheep virus disease yolk antibody prevents virus diseases through improving the antibody level of sheep antibodies on corresponding pathogen microorganisms causing the virus diseases. The feed additive of the invention is convenient to be fed to sheep with obvious control effect. The anti-sheep virus disease egg yolk antibody injection is provided with faster efficacy in the onset of the sheep. The anti-sheep virus disease yolk antibody feed additive, the injection and the preparing method thereof has the advantages of obvious efficacy, low cost, fast effect, no side effects, no drug-resistant strain which can be produced, simple preparation, wide raw material resource, convenient use and easy application.

The invention discloses a kind of bluetongue virus (BTV) NS2 protein monoclonal antibody BTV-4D4, a B cell epitope recognized thereby and applications. BALB / c mice are subjected to totivirus immunization by utilization of bluetongue virus serotype 8 type (BTV8), splenic lymphocytes of the mice after immunization and SP2 / 0 cells are fused. A hybridoma cell strain BTV-4D4 secreting anti-BTV-NS2 protein monoclonal antibodies steadily is obtained after screening with BTV 8 as a coating antigen through an indirect ELISA method. The monoclonal antibodies secreted from the hybridoma cell strain and the BTV-NS2 protein are subjected to a specific reaction. The B cell epitope recognized by BTV-4D4 is 149RSNYDV154 after identification by utilization of a phage display technology. The monoclonal antibody and the BTV-NS2 protein B cell epitope recognized by the monoclonal antibody can be used for diagnosis or prevention of bluetongue virus infection.

The invention discloses a preparation method for a bluetongue virus core sample grain and escherichia coli for simultaneously expressing bluetongue virus vp3 and vp7 proteins. The method comprises thefollowing steps: inserting bluetongue virus vp3 and vp7 genes into a dual-expression vector pETDuet-1, thereby constructing a pronucleus dual-expression plasmid pETDuet-VP3-VP7; converting into expression type escherichia coli BL21(DE3), thereby acquiring an expression bacterium containing the pronucleus dual-expression plasmid pETDuet-VP3-VP7; performing ultrasonic breaking and sucrose gradientcentrifugation after self-induced expression, thereby acquiring the bluetongue virus core sample grain. According to the invention, high-volume high-purity expression for the bluetongue virus core sample grain with excellent immunological competence is realized in the manner of prokaryotic expression.

The invention relates to the biotechnology field. A fast-detection test trip of a bluetongue virus (BTV) comprises a sample absorption pad 1, a nitrocellulose membrane 3, an absorbent pad 6 and a baseplate 7; wherein, the sample absorption pad, the nitrocellulose membrane and the absorbent pad are fixed on the baseplate by arrangement in sequence; the sample absorption pad is a porous fiber which is linearly arranged and fixed on an impermeable material; the sample absorption pad is loaded with a colloidal gold pad which is a conjugate of a monoclonal antibody of a BTV VP7 protein and the colloidal gold; a detection line 4 is positioned on the nitrocellulose membrane and a polyclonal antibody of the BTV is coated or sprayed on the detection line; a control line 5 is positioned on the nitrocellulose membrane and a protein A is coated or sprayed on the control line. The test strip of the invention features strong specificity, high sensitivity and fast detection, can show results in 10 minutes and is especially suitable for port quarantine, field and on-site quarantine. The test strip can detect the BTV antigen directly and rapidly, without special equipments or technicists.

The invention describes the preparation of an inactivated bluetongue virus (BTV) composition, and in particular a vaccine, using an attenuated live BTV vaccine strain as masterseed. The vaccine can be administered to an animal to prevent bluetongue disease by eliciting an immune response against the bluetongue virus serotype(s) included in the composition.

The invention discloses a multi-RT (reverse transcription)-PCR (polymerase chain reaction) reagent kit for genotyping identification of BTV (bluetongue virus) genotypes 2, 3, 4, 7 and 12 and a detection method of the BTV genotypes 2, 3, 4, 7 and 12, which are used for simultaneous identification and detection of the BTV genotypes 2, 3, 4, 7 and 12 through a single tube. According to different genotype virus VP2 gene sequence conservation areas of bluetongue, 5 pairs of PCR specific primers are designed in the method, a pair of non-biologically derived universal primers are synthesized by utilizing a GeXP principle, the universal primers are respectively added to the upstream and downstream of each pair of specific primers to form 5 pairs of specific chimeric primers, the specific primers are used for performing inverse transcription, and multi-PCR is constructed by combining the universal primers with the specific chimeric primers. By utilizing an optimized multi-PCR system and condition, one or more types of 5 genotypes, i.e., BTV genotypes 2, 3, 4, 7 and 12 can be simultaneously identified and detected through the single tube, other types of PTV, PPRV (peste des petits ruminantsvirus) and FMDV (foot-and-mouth disease virus) nucleic acids are not subjected to specific amplification, and the lowest detection concentration can reach a pg level. According to the method constructed by the invention, the sensitivity is high, the specificity is strong, the time and labor are saved, and the result is easy to observe.

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 provides a primer and a method for detecting Peste des petits ruminants virus and bluetongue virus. The primer combination designed and screened by the present invention finally successfully realizes the simple, rapid and accurate identification of Peste des petits ruminants virus (PPRV) and bluetongue virus in the same reaction tube for the first time through multiple fluorescent RT-LAMP detection methods (BTV). The multiple fluorescent RT-LAMP primers and method for detecting Peste des petits ruminants virus (PPRV) and bluetongue virus (BTV) provided by the present invention have good specificity, high sensitivity and low interference, and can detect at least 100 Mix template copies / reactions and effectively suppress false positive results. The primers and the method provided by the present invention can intuitively and accurately judge the results of the different colors displayed by the fluorescent groups, and only need a water bath to complete the amplification. The cost is low and the operation is convenient. It is a simple, fast, low-cost A cost-effective diagnostic method for large-scale epidemiological investigations of BTV and PPRV.

The invention discloses a 4-type bluetongue virus (BTV) VP2 protein neutralizing B cell epitope polypeptide and its application. The BTV VP2 protein epitope amino acids are screened by a phage display technology. After three rounds of screening, a sequencing process is carried out. Through analysis and comparison, a sequencing result shows that the common short peptide sequence is 160NH161. Experiment results show that 160NH161 can specifically react with the supernatant and ascites of 4A-1G7 and has good reaction specificity to 1 to 24-type BTV positive standard serum. The 4-type BTV VP2 protein neutralizing B cell epitope polypeptide provides a novel technical process for 4-type BTV clinical diagnosis and lays a basis for later building of a BTV-type specific identification and diagnosis method and preparation of a 4-type BTV related epitope vaccine.

The invention discloses a duplex fluorescent RT-LAMP detection group and kit for visually identifying foot-and-mouth disease virus and Bluetongue virus and an application of the detection group and the kit. The duplex fluorescent RT-LAMP detection group comprises two groups of specific primers and probes, the first group of specific primers and probes comprise FMDV-F3, FMDV-B3, FMDV-FIB(F1c-F2), FMDV-BIP(B1c-B2) and FMDV-probes, the second group of specific primers and probes comprise BTV-F3, BTV-B3, BTV-FIB(F1c-F2), BTV-BIP(B1c-B2) and BTV-probes, and the two groups of specific primers and probes are as shown in a sequence table SEQ ID No.1 to SEQ ID No.10. The duplex fluorescent RT-LAMP detection group can identify and diagnose the foot-and-mouth disease virus and the Bluetongue virus inthe same reaction tube and has the advantages of good specificity, high sensibility, less pollution, convenience, rapidness and the like, a detection result can be directly observed by naked eyes andis judged according to colors of reaction products, and the detection group can be used for basic-level quarantine with poor conditions.