35 results about "Hematopoietic organ" patented technology

The invention discloses a method for preparing a rainbow trout IHN (Infectious Haematopoietic Necrosis) inactivated vaccine, which comprises the following steps: carrying out grinding, filtering and poison dipping processing on pancreas and livers of juvenile fish which is attacked, but is still alive, inoculating rainbow trout gonad (RTG) cells, carrying out blind passaging at 14 DEG C, keeping for 5 days when carrying out blind passaging on the tenth generation, and collecting cell poisonous fluid; inoculating chinook salmon embryonic (CHSE) cells, carrying out passaging at 14 DEG C, and raising the culture temperature by 1 DEG C when passaging for 5 generations each time until the culture temperature is raised to 20 DEG C; and adopting epithelioma papulosum cyprini (EPC) cells to continuously carry out passaging under the condition of 20 DEG C, passaging to the twelfth generation to obtain high-titer virus solution and carrying out inactivation to obtain the rainbow trout IHN inactivated vaccine. According to the preparation method, RTG-2, CHSE-214 and EPC cells are utilized to alternately culture rainbow trout IHN viruses at a specific environment temperature so as to obtain a high-titer IHNV (Infectious Hematopoietic Necrosis Virus) antigen and produce the inactivated vaccine; and the technical difficult problem that the high-titer IHNV antigen cannot be obtained through single cells is solved.

The invention provides infectious hematopoietic necrosis (IHN) nucleic acid vaccines for Chinese rainbow trout and application thereof. A preparation method of the IHN nucleic acid vaccines for Chinese rainbow trout includes the following steps that firstly, specific primers for amplifying genotype J IHN virus surface glycoprotein genes (Glycoprotein, G) are designed; secondly, IHN virus isolates are cultured with a virus sensitive cell line, and then virus genome RNA is extracted; thirdly, the virus genome RNA obtained in the second step is subjected to one-step RT-PCR amplification with the primers obtained in the first step; fourthly, a recovery product obtained in the third step is connected with a pMD19-T simple vector, a connection product is converted into DH5alpha competent cells, single colonies are picked out, plasmids are extracted for PCR identification after amplified culture, and plasmids identified to be correct are subjected to sequencing; fifthly, IHNV isolate glycoprotein genes identified to be correct in the fourth step are cloned to a eukaryotic expression vector pcDNA3.1 with BamH I and Xho I restriction enzyme cutting sites to establish the nucleic acid vaccines.

The invention discloses an application of IL-2 protein to preparation of an animal vaccine adjuvant. The amino acid sequence of the IL-2 protein is shown as sequence 2 in a sequence table. Experiments prove that compared with Oncorhynchus mykiss injected with IHNV (infectious hematopoietic necrosis virus) G (glycoprotein), the level of expression of IgT gene, Mx gene, Viperin gene, CD4 gene, CD8 gene and TNF-alpha gene of Oncorhynchus mykiss injected with the IHNV G and the IL-2 protein is increased substantially, the titer of a neutralizing antibody is increased remarkably, proliferation of lymphocytes is enhanced greatly, and the protective efficacy of the IHNV G is improved effectively. Therefore, the IL-2 protein has important application value in preparation of the animal vaccine adjuvant.

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 compound medicine and method for preventing and treating infectious hematopoietic organ necrosis of Masu salmon, belonging to the technical field of aquaculture; 15%~25%, wild chrysanthemum 12%~18%, neem leaves 14%~16%, Atractylodes macrocephala 7%~13%, dried ginger 5%~15%, Houttuynia cordata 3%~7%, licorice 2%~ 8%; specific control methods include aeration of breeding water, methionine iodine bath and disinfection of breeding water, and vitamin C, vitamin E and compound drugs need to be added to the diet. The invention has simple operation, does not affect the quality of aquatic products, and is widely applicable to the prevention and treatment of infectious hematopoietic necrosis of masu salmon.

The invention discloses application of IL-17A protein in preparing animal vaccine adjuvants. The IL-17A protein is protein shown as an amino acid sequence 2 in a sequence table. Experiments prove that compared with rainbow trout injected with infectious hematopoietic necrosis virus G protein, expression level of IgT gene, IgM gene, Mx gene, Viperin gene and CD8 gene of the rainbow trout injected with the infectious hematopoietic necrosis virus G protein and the IL-17A protein is improved remarkably, valence of neutralizing antibody is improved remarkably, proliferation of lymphocytes is remarkably enhanced, and protection force of the infectious hematopoietic necrosis virus G protein is effectively increased. Therefore, the IL-17A protein has important application value in preparing the animal vaccine adjuvants.

The invention discloses a method for increasing blood storage protein (SP) amount of silkworm larvae in an early stage. Positions corresponding to hematopoietic organs of the silkworm larvae in a target stage are exposed to through holes of a separation plate, and the silkworm larvae are stimulated by heavy ion beams or laser irradiation through the through holes. When the heavy ion beams stimulation is carried out, the dose is controlled at 20-40 Gy. When the laser irradiation stimulation is carried out, the irradiation time is 20-80 s. The method better solves the problem that the amount ofthe blood SP of the silkworm larvae is less in the early fifth instar. The left and right sides of second and third thoracic segments of the silkworm larvae (where the hematopoietic organs exist) arephysically stimulated to induce the mass production of the SP in the early fifth instar of the silkworm. The induction treatment can increase the blood SP-1 content by 15-21% and the blood SP-2 content by 14-17%.

The invention relates to a nucleic acid vaccine for infectious hemopoietic necrosis of Chinese rainbow trouts and application of the nucleic acid vaccine. A preparation method of the nucleic acid vaccine includes (1), designing and amplifying specific primer pairs of surface glycoprotein of genetype-J infectious hemopoietic necrosis viruses; (2), culturing IHN (infectious hemopoietic necrosis) virus isolates by virus sensitive cell lines, and extracting viral genome RNA; (3), performing one-step RT-PCR amplification on the viral genome RNA obtained in the step (2) by the primer pairs obtained in the step (1), and recycling RT-PCR amplified products after electrophoresis; (4), connecting the recycled products in the step (3) with pMD19-T simple carriers, converting connection products into DH5alpha competent cells, picking single colonies for amplification and culture, extracting plasmids, performing PCR verification, and sequencing the correct plasmids; (5), cloning open reading frame portions of the correct IHNV (infectious hemopoietic necrosis virus) isolate glycoprotein genes in the step (4) to eukaryotic expression vectors pcDNA 3.1 by the aid of BamH I and Xho I digestion points to create the nucleic acid vaccine.

The invention discloses primers for detecting the epidemic haematopoietic necrosis virus with a pyrosequencing technology, a kit and a detection method, and particularly relates to a group of primers for detecting the epidemic haematopoietic necrosis virus with the pyrosequencing technology. The primers comprise an amplification primer pair shown in sequence tables SEQ ID No.2 and SEQ ID No.3 as well as a sequencing primer shown in the sequence table SEQ ID No.4. The invention further discloses the detection kit containing those primers. The pyrosequencing technology is used for detecting the epidemic haematopoietic necrosis virus and has the advantages of high flux, rapidness and accuracy, short DNA sequence synthesis can be performed accurately, the sequencing process takes a short time, and construction of the standard operation process is facilitated.