54 results about "Kidney necrosis" patented technology

The invention discloses a method for detecting infectious spleen and kidney necrosis viruses by using hyper-branched rolling circle amplification, which is characterized by comprising the following steps of: (1) designing one locking type probe and a pair of universal primers for the connection sequence of the locking type probe; (2) making a connecting reaction system of the locking type probe for connecting reaction; and (3) making an HRCA (Hyper-branched Rolling Cycle Amplification) reaction system for HRCA reaction, and finally detecting the products of the HRCA reaction. Compared with the method for detecting the infectious spleen and kidney necrosis viruses by using PCR (Polymerase Chain Reaction) in the prior art, the method has the advantages of higher sensitivity, specificity and convenience, can be applied to actual production sites and is beneficial to detecting and controlling the cross infection of the infectious spleen and kidney necrosis viruses in the cultivation of aquatic animals.

The invention discloses a proliferation method of a siniperca chuatsi infectious spleen and kidney necrosis virus (ISKNV). According to the proliferation method, synchronous virus inoculation is adopted, the virus and a siniperca chuatsi brain tissue cell line (CPB) are in a suspended state, and the mutual contact surface area between the virus and the siniperca chuatsi brain tissue cell line is large so that the virus enters cells through more space receptors; and the virus infects the cells while the cells differentiate and proliferate, thus obtaining the high content ISKNV. After the CPB is cultured for 30 hours to 60 hours, the CPB is transferred to 5%-8% v / v of fetal calf serum culture medium, and the ISKNV is synchronously inoculated at the MOI (Multiplicity of Infection) value of 0.85-2.3, thus obtaining a low-cost ISKNV proliferation method. After the CPB is cultured for 20 hours to 40 hours, the CPB is transferred to 8%-10% v / v of fetal calf serum culture medium, and the ISKNV is synchronously inoculated at the MOI (Multiplicity of Infection) value of 30-45, thus obtaining a short ISKNV proliferation method. Cultivation methods can be selected according to existing conditions. The proliferation method provides theoretical basis for production of low-cost viruses.

The invention relates to a marine fish embryo cell culture technology, in particular to an establishing method of a myosatellite cell system of bastard halibut during an embryonic period. The establishing method comprises the following steps: carrying out in-vitro separation on an embryo during a bastard halibut kirschner capsule formation period, removing an egg membrane, dispersing cells, and carrying out primary culture and secondary culture, thus establishing the myosatellite cell system during the embryonic period. The form of the myosatellite cell system, established by the invention, ofthe bastard halibut is fusiform or spindle-shaped mononuclear cells; myofiber can be differentiated to form by culturing for 5 days or above within a single generation or carrying out induction of anexogenous factor (horse serum). The myosatellite cell system is capable of supplying a large amount of myosatellite cells, and GFD (Green Fluorescent Protein) transfection and cynoglossus semilaevisspleen and kidney necrosis virus challenge assay detection verify that the myosatellite cells can be normally transfected or infected, so that the myosatellite cell system not only can be directly used for researching functional genes related to muscle growth and development of fishes and providing research materials for exploring a molecular mechanism for induction, proliferation and differentiation of muscle cells of the fishes, but also is capable of providing a platform for improved research and application of muscle characters in bastard halibut breeding.

The invention discloses a method for producing a siniperca chuatsi infectious spleen and kidney necrosis virus and a siniperca chuatsi rhabdovirus through microcarrier suspension culturing CPB cells. The method comprises the steps of selecting the CPB cells as a cell line for making vaccines; sequentially carrying out step-by-step magnifying microcarrier suspension culture on the CPB cells through stirring bottles and bioreactors, so that the CPB cells are used as hosts of virus infection; finally obtaining the siniperca chuatsi infectious spleen and kidney necrosis virus bulk and the siniperca chuatsi rhabdovirus bulk. The prepared ISKNV potency reaches up to 8.34LgTCID50 / mL, and the SCRV potency reaches up to 10.25LgTCID50 / mL. A microcarrier suspension culture process adopted by the invention is simple to operate, less in pollution probability, small in labor force compared with a roller bottle culture method, and lower in production cost.

The invention discloses a triple PCR primer group for rapidly detecting three fish viruses and application thereof, and relates to the technical field of gene detection. The invention discloses a triple PCR primer group for rapidly detecting three fish viruses. The triple PCR primer group comprises an infectious spleen and kidney necrosis virus ISKNV primer, a mandarin fish frog virus SCRIV primerand a mandarin fish rhabdovirus SCRV primer, the triple PCR primer group has high sensitivity, the detection time is 2-3 hours earlier than that of conventional PCR, the efficiency of clinical diagnosis is greatly improved, richer reference bases can be provided for epidemiological investigation of ISKNV, SCRIV and SCRV infection, and the triple PCR primer group has important significance for subsequent research and prevention and treatment.