39 results about "Cyprinid herpesvirus" patented technology

The present invention discloses a cyprinid herpesvirus II-sensitive brain tissue cell line of Carassius auratus gibelio and establishing method and use thereof. The cell line is deposited in China Center for Type Culture Collection under an accession number of CCTCC No: C2013179. The brain tissue cell line of Carassius auratus gibelio is in good growth state and sensitive to CyHV-2 that is presently hardly cultured with ordinary fish cell lines. After six passages of CyHV-2 in GiCB cells, viral nucleic acid can still be detected and a cytopathic effect is stable. When ultrathin microscopic sections are prepared from cells having the cytopathic effect, considerable mature CyHV-2 virions and their replication process can be observed in GiCB cells. The construction method of the brain tissue cell line of Carassius auratus gibelio of the present invention has high repeatability, scientific and reasonable conditions.

The invention discloses an immunity index for indicating cyprinid herpesvirus resistance and relates to an immunity index for indicating cyprinid virus resistance. The immunity index for indicating cyprinid herpesvirus resistance, disclosed by the invention, includes activity of alkaline phosphatase (AKP). According to the invention, the immunity index for indicating cyprinid herpesvirus resistance is found, and the activity of the infected immunity index molecule for indicating cyprinid herpesvirus resistance is consistent with the survival rate of the infected carp. Therefore, the immunity index for indicating cyprinid herpesvirus resistance, disclosed by the invention, can accurately reflect the disease resistance.

The invention belongs to the technical field of vaccine preparation, and particularly provides a cyprinid herpesvirus 2 attenuated strain and an application thereof. The cyprinid herpesvirus 2 provided by the invention is a cyprinid herpesvirus 2 DX2019 strain, and the preservation number is CCTCC NO:V201980. A vaccine prepared by using the strain can prevent diseases caused by the cyprinid herpesvirus 2, has an immune protection effect of 92 percent or above, does not generate a stress reaction to organisms, and can be applied to prevention of haematopoietic organ necrosis of crucian carps, so that the incidence rate of haematopoietic organ necrosis of the crucian carps in a crucian carp culture pond is reduced, and the survival rate of the crucian carps is improved.

The invention relates to a cyprinid herpesvirus II DNA vaccine based on a baculovirus vector as well as a building method and application thereof. Codons of regional sequences 1-186, 993-1197, 603-783and 85-186 nt of cyprinid herpesvirus II ORF72, ORF66, ORF81 and ORF82 are optimized; then, ORF72-ORF66-ORF81-ORF82 fusion sequences are chemically synthesized; next, the promoter control is performed; the clone into the baculovirus transfer vector pFSATBacTMDual is performed to build recombinant plasmids pFSATBacTMDual-FA-D4ORF; after the plasmids are used for converting DH10 / Bac competent cells, Bacmid-FA-D4ORF is obtained; after the Bacmid-FA-D4ORF DNA is used for transfecting silkworm culture cells, BmNPV-FA-D4ORF is obtained and is the cyprinid herpesvirus II DNA vaccine based on the baculovirus vector. The DNA vaccine can realize the immune on carassius auratus gibelio by using an injection or oral administration or soaking method; the gill bleeding diseases of the carassius auratusgibelio can be reduced.

The invention relates to a cyprinid herpesvirus II type sandwich ELISA detection kit. The kit mainly comprises a cyprinid herpesvirus II type ORF90 purified monoclonal antibody solution, an enzyme-labeled ORF90 polyclonal body solution, a diluent, a TMB substrate solution, a stop solution, cyprinid herpesvirus II type positive control, negative control, coating liquid and a 96-pore enzyme-labeled plate. The invention also relates to a detection method. The kit and the method, provided by the invention, can be applied to detection of latent infected fish virus without clinical symptom and can further be applied to research on the change rule of the cyprinid herpesvirus II type in the fish body, so that basis is provided for prevention and control of the virus. The method provided by the invention does not have specific reaction with other common viruses of crucian and goldfish, and can be applied to diagnosis of epidemic disease in domestic farms as well as exit and entry fish quarantine.

The invention relates to a Carassius auratus gibelio anti-Cyprinid herpesvirus 2 (CyHV-2) oral recombinant Bacillus subtilis spore vaccine and a preparation method thereof. The vaccine is prepared by coating recombinant Bacillus subtilis spore CyHV-2 having a surface showing a CyHV-2 immune protective antigen with a Carassius auratus gibelio granular feed; and the above spore is a spore with germination defects and no antibiotic-resistant gene markers. The preparation method comprises the following steps: constructing the recombinant Bacillus subtilis spore CyHV-2 having germination defects, having no antibiotic resistance gene markers and having a surface showing the CyHV-2 immune protective antigen, and preparing the Carassius auratus gibelio granular feed coated CyHV-2 oral recombinant Bacillus subtilis spore vaccine used for preventing and treating CyHV-2 induced Carassius auratus gibelio hemorrhage. The oral recombinant Bacillus subtilis spore vaccine having germination defects, having no antibiotic resistance gene markers and having a surface showing the CyHV-2 immune protective antigen, prepared in the invention, has the characteristics of safety and high efficiency.

The invention discloses a CyHV-2 (Cyprinid Herpesvirus 2) monoclonal antibody, a hybridoma cell and an application of the CyHV-2 monoclonal antibody. The CyHV-2 monoclonal antibody disclosed by the invention is secreted from a hybridoma cell GFHNV-6G7 with the collection number of CCTCC No. C2013114 or hybridoma cell GFHNV-3B5 with the collection number of CCTCC No. C2013115. The CyHV-2 monoclonal antibody disclosed by the invention has good specificity and potency; by producing rapid immunodetection test paper from the CyHV-2 monoclonal antibody, the speed and efficiency of detection can be increased, and field detection on CyHV-2 can be realized. The CyHV-2 monoclonal antibody disclosed by the invention makes up the blank in immunodetection on the CyHV-2 and lays a foundation for immunological research on the CyHV-2.

The invention discloses the application of ferrous ions in the prevention and treatment of cyprinid herpesvirus 2 (CyHV2) infection. The establishment of experimental animal infection models shows that ferrous chloride can significantly protect crucian carp from CyHV2 infection. Combined with the results of qPCR experiments, it was found that the transcription level of virus-related genes was significantly inhibited after ferrous chloride treatment. Therefore, the compound containing ferrous ions can be used as a drug for preventing and treating CyHV2, and has the advantages of non-toxic and safe dosage and good protective effect.