34 results about "Koi herpesvirus" patented technology

The invention discloses a dual real-time fluorescent quantitative PCR detection method for carp edema virus and koi herpes virus. The method includes the following steps: first, synthesizing primers and TaqMan probes: optimizing the primer sequence according to the CEV P4a gene sequence, The first base of the upstream primer was changed to a degenerate base W, and the first base of the downstream primer was changed to a degenerate base R; according to the conserved sequence of the KHV ORF7 gene, a pair of specific primers and a TaqMan probe were designed; Use FAM and VIC as the probe reporter group, and BHQ1 as the probe quencher group; second, determine the reaction system and conditions: use 20 μL reaction system, 2×Probe PCR Master Mix 10 μL, upstream and downstream primers and probes The final primer concentration was between 0.2 μmol / L and 0.8 μmol / L, QN ROX reference dye 0.1 μL, template 2.5 μL, and DEPC water to make up 20 μL; the reaction program was: 95°C pre-denaturation for 2 min, 1 cycle; 95°C for 5 s, Anneal at 50-60°C for 31s, 40 cycles.

The present invention provides a DNA vaccine for carps for inducing protective immunity against Koi herpesvirus (KHV). The DNA vaccine comprises a DNA comprising a nucleotide sequence encoding an immunogenic polypeptide against Koi herpesvirus (KHV) of carps, or an expression vector comprising the DNA as an active ingredient.

The invention discloses a KHV (koi herpes virus) CPA (cross priming amplification) detection primer and an application. A KHV CPA detection kit comprises following ingredients: 10*ThermoPol Reaction Buffer, Bst DNA polymerase, dNTPs, cross primers CPF and CPR, detection primers DF and DR, stripping primers DPF and DPR, MgSO4, Betaine and a nucleic acid test strip. The KHV CPA detection primer has the characteristics of simplicity, convenience, rapidness and high specificity and sensitivity and is objective and direct in determination of results, low in cost, convenient to use and very safe to people and the environment. The KHV CPA detection primer not only can be used in a special laboratory, but also can be applied to rapid onsite detection in the field, only one metal bath or water bath kettle is required, and a KHV in a sample can be accurately detected within 1.5 h.

The invention relates to the technical field of aquatic antiviral microecological preparations, in particular to a bacillus licheniformis YFI‑2 and its application in the preparation of freshwater cultured animal virus disease drugs. The preservation number of the Bacillus licheniformis of the present invention is CCTCC NO: M2019652. The fermentation supernatant of Bacillus licheniformis YFI‑2 in the present invention can reduce grass carp reovirus (GCRV), koi herpes virus (KHV), carp herpes virus type 2 (CyHV‑2), channel catfish reo Viruses (CCRV) and giant salamander iridescent virus (GSIV) have infective activity on host cells, inhibit virus invasion and infection of cells, and have no toxic side effects on cells, and can be used to prevent and treat viral diseases in freshwater farmed animals.

The invention discloses a method for constructing a koi brain cell line and its application. The koi brain tissue is first put into digestion solution A containing collagenase and trypsin for digestion, and the cells are collected, and then added into a digestion solution containing EDTA‑4Na. Digest in solution B, collect the digested cells for primary culture and subculture to obtain the koi brain cell line; the present invention aims at the characteristics of koi brain tissue cells, and uses collagenase and trypsin joint digestion method to separate brain tissue The cells were cultured in the first generation, and then the koi brain cell line was successfully constructed, which has been continuously passed on for more than 100 generations. It can provide a large number of koi brain-derived cells, and the cells can maintain a good growth state, which can be cryopreserved . The koi brain cell line of the present invention is sensitive to the koi herpes virus, and cytopathy occurs after inoculation, and multiplied virus particles can be found by electron microscope observation. The cell line of the present invention can be directly applied to the research of pathogenic characteristics and the development of vaccines.

The invention belongs to the technical field of biology and discloses a primer set for the detection of a koi herpesvirus Sph gene and the application of the primer set. The premier set for the detection of the koi herpesvirus Sph gene comprises a forward outer primer having the nucleotide sequence shown in SEQ ID No.1, a backward outer primer having the nucleotide sequence shown in SEQ ID No.2, a forward inner primer having the nucleotide sequence shown in SEQ ID No.3 and a backward inner primer having the nucleotide sequence shown in SEQ ID No.4. The primer set provided by the invention is high in specificity and sensitivity, and suitable for amplification of koi herpesvirus Sph gene through an LAMP (Loop-Mediated Isothermal Amplification) method, and can be used for preparation of a reagent for detection of the koi herpesvirus Sph gene.

The invention discloses a preparation method and application of a monoclonal antibody against koi herpesvirus (KHV). The prepared monoclonal antibody against KHV is secreted by a mouse hybridoma cellstrain of which the preservation number is CCTCC NO: C201887, and the titer of the monoclonal antibody is 1:(1.28 is multiplied by 105); and the hybridoma cell strain has high activity, can recognizethe structural protein of the KHV, and has strong specificity. On the basis, a sandwich ELISA detection method for detecting the KHV is established, and the minimum detection concentration is 3.923ng / mL. The monoclonal antibody is utilized for establishing an indirect sandwich ELISA kit for detecting the KHV, and the kit can be used for large-scale detection of the KHV and has wide application prospects.

The invention relates to cloning, sequencing, analysis and functional study of a biosynthetic gene cluster of a natural product pentostatin which is produced from streptomyces antibioticus and is used for treating lymphocytic leukemia and a natural product arabinofuranosyladenine for resisting Koi herpes virus and varicella-zoster virus and application of the biosynthetic gene cluster. Biosynthetic genes of the two compounds are included in the same gene cluster and are independent from each other. The whole gene cluster comprises ten genes, namely three genes related to pentostatin synthesis, five genes related to arabinofuranosyladenine synthesis and two genes related to transportation regulation of the pentostatin and arabinofuranosyladenine. According to genetic manipulations of the previous biosynthetic genes, biosynthesis of the pentostatin or arabinofuranosyladenine can be blocked or improved. The genes provided by the invention and proteins thereof can be used for genetic engineering, protein expression, enzymic catalytic reaction and the like of the compounds, and can be further used for searching and discovering compounds or genes and proteins which can be used for medicines, industry or agriculture.