21 results about "Virus inoculation" patented technology

The application discloses an isolation method of bovine epidemic fever virus and the bovine epidemic fever virus obtained by the method and application of the bovine epidemic fever virus, and belongs to the technical field of bovine epidemic fever virus isolation. The application aims to provide an efficient and rapid method for isolating bovine epidemic fever virus. The application provides the isolation method of the bovine epidemic fever virus, wherein the anticoagulated blood sample of a bovine is inoculated on KC cells for culture, the virus is harvested, the BHK-21 cells are inoculated to grow into a monolayer, and the first generation of passage cells with cytopathic effect is obtained by passage. The virus strain can also be used for screening, preparation and detection of bovine epidemic fever virus antibodies, and for preparation of an animal model infected by the bovine epidemic fever virus.

The invention discloses a cat kidney suspension culture cell line suitable for serum-free suspension culture and application of the cat kidney suspension culture cell line. The name of the cell line is F81-B4, and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.46350. After 50 generations of continuous subculture, the cells still can maintain the original proliferation level and cellular morphology, the cell number can reach 9.0 * 10 < 6 > / mL after the cells are subcultured at the density of 1.0 * 10 < 6 > / mL for three days, the multiplication time is 22-26 hours, and the cell proliferation speed is equivalent to that in a shake flask when the cells are amplified to a 10L bioreactor for culture. The cell line is highly susceptible to feline parvovirus, feline calicivirus, feline herpes virus, canine parvovirus and canine distemper virus, the virus content of a virus solution harvested 72 h after FPV virus inoculation can reach 107.5 TCID50 / mL, the virus content of a virus solution harvested 24 h after FCV virus inoculation can reach 1010.8 TCID50 / mL, the virus content of a virus solution harvested 48 h after FHV-1 virus inoculation can reach 107.5 TCID50 / mL, compared with an existing F81 adherent cell production process, the production process has the advantages that the production efficiency is high, and the production cost is low. The method has the characteristics of rapid lesion, short virus collection time and high titer. The cell matrix is an ideal cell matrix which can be used for culturing the three viruses.

The utility model relates to the technical field of plant virus inoculation devices, in particular to a plant virus friction virus receiving device which comprises a liquid taking box and a friction virus receiving assembly. The liquid taking box is detachably connected with the friction poison receiving assembly; the friction poison receiving assembly comprises a bottom supporting box, the bottom supporting box is movably connected with a poison receiving box, the bottom supporting box is detachably connected with a liquid taking box, and the liquid taking box and the poison receiving box are oppositely arranged; a plurality of bristles are arranged on the inner wall of the poison receiving box, and the length of the bristles is equal to the depth of the liquid taking box; a movable bottom plate is arranged in the poison receiving box, the movable bottom plate can move towards or away from the liquid taking box, and the bristles movably penetrate through the movable bottom plate. According to the friction virus receiving device provided by the utility model, the friction stress of the blades can be strictly controlled, and the stress of the whole blades is uniform, so that the condition of insufficient friction or excessive force during inoculation is avoided. The device is suitable for laboratory novice, and can reduce the number of times of experiment groping and failure.

The utility model discloses a cell tolerance testing device, which belongs to the technical field of cell culture and comprises a shell, a switch door is arranged on one side of the shell, a liquid injection device is fixedly connected onto the shell, a receiving device and a rotating device are mounted in the shell, the liquid injection device comprises a first motor and an injection cylinder, and the first motor is connected with the rotating device. The output end of the first motor is fixedly connected with a piston, and the piston is arranged in the injection cylinder. According to the utility model, when virus inoculation liquid needs to be inoculated into cell liquid, the virus liquid only needs to be injected through the one-way water inlet valve, in the operation process, the first motor is started, the motor drives the piston to axially move along the inner part of the injection cylinder and applies certain pressure to the virus inoculation liquid in the injection cylinder, and the virus inoculation liquid is inoculated into the cell liquid under the action of the pressure; and the virus inoculation liquid is discharged through the liquid outlet nozzle, then flows into the flow guide nozzle and is finally injected into the corresponding measuring cup below, so that pollution-free and quantitative injection of the virus inoculation liquid is realized.

The application discloses a preparation method of a tetravalent influenza virus split vaccine and belongs to the technical field of vaccines. The preparation method of the tetravalent influenza virus split vaccine comprises the following steps: pre-incubation of a chicken embryo, pre-embryo inspection, virus inoculation and culture, post-embryo inspection, cold embryo, virus harvesting, ultrafiltration clarification, zone centrifugation, desugaring, chromatography, virus splitting, virus inactivation, ultrafiltration liquid replacement, sterilization filtration and preparation of a tetravalent influenza virus split vaccine product. Compared with the prior art, the technical means adopted in the application effectively reduces the content of free formaldehyde and improves the antibody level of the vaccine.

The invention relates to the fields of molecular biology and virology, and provides a method for preparing a specific MHC haplotype-derived cell-binding virus. The method comprises the following steps: firstly, obtaining chick embryos of different haplotypes, separating CEF cells, transiently expressing exogenous fluorescent protein through electrotransfection, then carrying out virus inoculation, carrying out flow sorting to obtain a fluorescent positive cell population carrying MDV, and carrying out quantitative analysis on the cell population to obtain the specific MHC haplotype chicken Marek's Disease Virus (MDV), thereby obtaining the specific MHC haplotype chicken Marek's Disease Virus Marek's Disease Virus Marek's Disease Virus Marek's Disease Virus. And carrying out in-vitro propagation to obtain the purified MDV with a specific haplotype source. The purified virus can provide a test material for related research of haplotype on virus infection, and can be widely applied to in-vitro purification of different cell binding viruses and related research of specific MHC molecule presenting virus peptides. The method disclosed by the invention is beneficial to carrying out related research aiming at the host immune response of the virus and analyzing the hereditary basis of haplotype to MDV differential resistance, and is expected to be applied to preparation of vaccine virus.

The invention relates to the technical field of biological medicine, in particular to a construction method and application of a respiratory virus infected alveolar organ model. Comprising the following steps: infecting alveolar cell source single cells by using a virus inoculum, coating the infected alveolar cell source single cells with matrigel, and inoculating the infected alveolar cell source single cells into a first culture medium for three-dimensional culture to obtain the respiratory virus infected alveolar organ model. According to the invention, alveolar cells are creatively applied to model characterization of respiratory virus infection, and particularly, alveolar cell source single cells are subjected to infection treatment by adopting influenza A H1N1 virus, so that the constructed model can well simulate and characterize the infection process of influenza A virus in human lung organs; the method provides an experimental basis for drug research and development (such as high-throughput screening of anti-influenza virus drugs), viral disease pathogenesis research, research on defense mechanisms of naturally occurring virus infection or virus infection caused by experiments and the like, and has important guiding significance.

The invention relates to a method for improving PCV2 recombinant baculovirus protein expression, and belongs to the technical field of recombinant virus protein preparation. In order to solve the problem of low expression quantity of the existing PCV2 recombinant virus protein, the invention provides a method for efficiently expressing the PCV2 recombinant virus protein, the method comprises the step of carrying out H5 cell suspension culture for preparation, and we find that when glucose is lower than 5g / L in the culture process, glucose is added to 5-8g / L, glutamine is supplemented to 3-5mM, and beta-mercaptoethanol is added to 0.01-0.1 mM, the expression quantity of the PCV2 recombinant virus protein is increased, and the expression quantity of the PCV2 recombinant virus protein is increased. The expression quantity of the target protein can be obviously increased, and the expression quantity reaches 240 g / L or above. The method is simple in process, condition parameters are easy to control, cell culture is extremely easy to amplify, and the effects of cell culture, virus passage and target protein expression can be achieved without changing liquid in the cell culture period after virus inoculation.

The application relates to application of a dihydropyridine compound or a salt thereof in preparation of a medicine for preventing and / or treating HPV infection, and belongs to the technical field of medicines and pharmaceutical preparations. The application provides application of the dihydropyridine compound or the salt thereof in preparation of the medicine for preventing and / or treating HPV infection, wherein the dihydropyridine compound is selected from one or more of nifedipine, amlodipine, lercanidipine, nimodipine, nitrendipine, nisoldipine, felodipine, benidipine, lacidipine and azelnidipine. The application scheme first reports the application of the dihydropyridine compound or the salt thereof in prevention and / or treatment of HPV, and in-vitro cell experiments are carried out through azelnidipine in the dihydropyridine compound, and further, the inhibiting effect of the dihydropyridine compound on HPV virus is verified through medicine administration of a mouse intradermal virus inoculation model and medicine administration of a mouse vaginal virus inoculation model.

The invention belongs to the technical field of plant gene function research, and particularly relates to a method for inducing endogenous gene silencing of common tobacco (Nicotiana tabacum) by using a tobacco embrittlement virus. In order to solve the problems of unstable silencing efficiency, poor systematic silencing effect and the like of the existing TRV-VIGS technology in common tobacco, various key parameters about tobacco varieties, virus inoculation methods and plant culture conditions in the implementation process of the TRV-VIGS technology are determined, and the problems of common tobacco are effectively solved by adopting the method provided by the invention. The endogenous NtPDS gene of common tobacco is successfully silenced, and the silencing efficiency is as high as 99.02%.

The invention discloses a negative pressure fixation and micro-injection combined cotton virus-induced gene silencing inoculation method. The method comprises the following steps: (1) constructing a virus-induced gene silencing vector; (2) preparing an agrobacterium suspension; (3) fixing under negative pressure; (4) performing micro-injection inoculation; and (5) subsequent culture: culturing the inoculated plant under the conditions that the temperature is 22-25 DEG C, the illumination period is 16h illumination / 8h darkness, the illumination intensity is 100-200 and the relative humidity is 60-80%, and observing the gene silencing phenotype after 7-14 days. The method solves the problems that a traditional virus-induced gene silencing inoculation method is complex in operation, low in success rate and large in leaf damage, has the advantages of being easy and convenient to operate, high in inoculation success rate, small in leaf damage, high in gene silencing efficiency and the like, and is suitable for cotton functional genomics research and gene function verification.

The application discloses a construction method of Aedes albopictus Aa01 cell line and susceptible cell model of Decapod iridovirus 1, and belongs to the technical field of virology and aquaculture disease prevention and control, wherein the Aedes albopictus Aa01 cell line is preserved in the China Center for Type Culture Collection of Wuhan University, and the preservation number is CCTCC NO: C202648. The method for constructing the DIV1 susceptible cell model based on the above cell line comprises the following steps: culturing the Aedes albopictus Aa01 cell line, and subculturing or using for virus inoculation when the density reaches 80%-90%; inoculating the DIV1 into the Aedes albopictus Aa01 cell line, removing the virus liquid after virus adsorption, and adding cell maintenance liquid for continuous culture; and observing the cytopathic effect CPE, collecting the cell culture when the CPE reaches 70%-80%, and obtaining the amplified DIV1 virus liquid. The DIV1 susceptible cell model constructed by the method can solve the problem that there is currently a lack of stable and continuously subcultured DIV1 in vitro culture.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to application of an RNA interference fragment of a targeted Pt15401 gene and a related biological material of the RNA interference fragment to improvement of leaf rust resistance of crops. The invention finds that Pt15401 is a typical secretory protein, and heterologous expression in tobacco finds that Pt15401 can inhibit Bax-induced cell necrosis and exert a cytotoxic function. Furthermore, the Pt15401 gene is interfered based on a host-induced gene silencing (HIGS) technology to obtain an instantaneous silent crop plant, and virus inoculation verification on the instantaneous silent crop plant finds that the leaf rust resistance of the instantaneous silent crop plant is remarkably enhanced, and the leaf rust resistance of the instantaneous silent crop plant is remarkably improved. Furthermore, the application of the RNA interference fragment of the targeted Pt15401 gene and related biological materials in the aspect of improving the leaf rust resistance of the crops is proved, and gene resources and technical support are provided for subsequent breeding of the leaf rust resistant crops.

The invention discloses a method for preparing an rHP-PRRSV-NADC30 Nsp230 + 131 recombinant strain and an application of the rHP-PRRSV-NADC30 Nsp230 + 131 recombinant strain. According to the present invention, by using the reverse genetic manipulation technology, the virus strain is successfully constructed and saved, and the inherent 30 deletion of HP-PRRSV (line 8.7) and the 131 deletion of NADC30-like (line 1.8) are simultaneously retained on the PRRSV Nsp2 protein of the virus strain; the modified virus is inoculated to Marc-145 cells for subculture, and experimental results show that the modified virus can induce cytopathy. The obtained recombinant strain shows a stable passage capability in a Marc-145 cell. In addition, clinical animal experiments further verify the safety and effectiveness of the recombinant strain. No obvious pathological reaction and abnormal clinical manifestation are observed in an experimental animal inoculated with the recombinant strain, which indicates that the recombinant strain has good biological safety. Meanwhile, experimental animals show that the recombinant strain can stimulate effective immune response, an effective immune protection effect is provided for multilineage PRRSV infection, and potential vaccine development prospects are shown.

The invention relates to the technical field of veterinary medicine and animal models, in particular to a modeling method for inducing broiler chicken immunosuppression and secondary bacterial infection. The method comprises the following steps: (a) starting to continuously feed broiler chickens with daily ration containing mycotoxin at the age of 6-10 days until molding is finished; (b) inoculating infectious bursal disease virus on the broiler chicken at the age of 12-16 days; and (c) inoculating avian pathogenic escherichia coli after inoculating the infectious bursal disease virus for a period of time. According to the modeling method, the clinical pathogenesis process is highly simulated, the mechanism fits the reality, the model success rate is high and stable, standardized operation is achieved, repeatability is good, simplicity, convenience and safety are achieved, and the biological safety risk of virulent strains is avoided; the application range is wide, and an excellent platform is provided for research and development; the method is low in cost and suitable for scientific research and large-scale drug screening popularization.

The invention discloses an inoculating and harvesting all-in-one machine, which comprises a disinfection device for disinfecting the upper surface of an embryo egg; the inoculation equipment comprises an egg jacking module, an inoculation module and a disinfection module, the inoculation module comprises an inoculation needle, and the disinfection module is used for disinfecting the inoculation needle; the harvesting equipment comprises a shell cutting module, a harvesting module and a detection module, the shell cutting module is used for cutting off the upper eggshell of the embryo egg, the detection module is used for detecting whether the embryo egg is qualified or not, the harvesting module comprises a plurality of harvesting heads, and the detection module is used for detecting the quality of the harvested liquid; and the caching equipment is used for caching the egg trays. The device has the beneficial effects that the processes of embryo egg conveying, embryo egg disinfection, egg jacking, punching, virus inoculation, embryo egg shell cutting, embryo egg screening and allantoic fluid harvesting are achieved, the processing efficiency is high, pollution is reduced, and the production quality is improved.

The application discloses a QMC influenza virus seed, a preparation method and application thereof, and belongs to the technical field of biological medicines.The preparation method of the QMC virus seed is virus passage adaptation and virus seed preparation which are synchronously performed, and comprises the following steps: inoculating chicken embryo source influenza virus on MDCK-XF06 cells, and obtaining recovered passage virus seed after culture; then inoculating the recovered passage virus seed on MDCK cells according to a mode of adjusting virus inoculation infection multiplicity from generation to generation, and obtaining P2 generation master seed batch virus seed and P3 generation working seed batch virus seed after culture.The QMC virus seed obtained through the preparation method has the following advantages: good virus activity, high virus titer, efficient production of a large amount of live virus, excellent genetic stability, ensured consistency and effectiveness of product quality, completely eliminated tumorigenicity risk, high safety, and the like, and lays a solid foundation for subsequent production of safe, efficient and stable vaccine products.

The application discloses an influenza virus subunit vaccine and a preparation method thereof, and relates to the field of biological medicines. Each dose contains H1N1, H3N2 and Victoria; the influenza virus subunit vaccine is prepared through virus inoculation, proliferation culture, allantoic fluid harvesting, clarification, inactivation, ultrafiltration concentration, lysis purification, mixing, sterilization filtration and subpackaging. The application significantly enhances the dendritic cell targeted uptake and rapid release of antigens through the design of mannose modified small liposomes and pH responsive channels; in combination with the precise controlled release of the graded lysis micelles, the antibody titer and immunological persistence are obviously improved, the single dose protection period is longer, and the defects of the traditional trivalent vaccine in the insufficient immunological intensity are effectively made up.

Impedance-based determination of metrics derived from Cytopathic Effects (CPE), such as Tissue Culture Infections Dose (TCID) values, is provided via determining a first plurality of cellular impedances (CI) corresponding to a plurality of cell samples while performing the impedance assay; inoculating the plurality of cell samples with a plurality of different dilutions of the virus such that a first and second subsets of the plurality of cell samples are respectively inoculated with first and second dilutions of a virus; determining, after inoculating the plurality of cell samples, a second plurality of CIs; determining, from the first plurality of CIs and the second plurality of CIs, cytopathic effects (CPE) experienced by the plurality of cell samples; calculating, a CPE-derived metric.

The invention relates to the technical field of bioengineering, in particular to a screening method for culturing O-type FMDV high-yield cell strains. A screening method for culturing the O-type FMDV high-yield cell strain is established by an indirect immunofluorescence labeling method, and the cell strain with strong infectivity and high sensitivity to the O-type FMDV is screened by simultaneously detecting whether the proportions of alpha v beta 1 and alpha v beta 6 positive receptors on the surface of a BHK-21 cell in population cells are greater than or equal to 25%. The screening method can accurately pre-judge the performance of the cell strain before virus inoculation, the screening direction is clear, and the result is reliable. According to the method, virus inoculation, virus collection and virus content detection are not needed, screening can be completed within 4 days, the period is greatly shortened, the cost and the biological safety risk are reduced, operation is easy and convenient, repeatability is good, and a stable and reliable cell base material is provided for production of efficient foot-and-mouth disease inactivated vaccines.

The invention discloses a preparation method of a tetravalent influenza virus split vaccine, and belongs to the technical field of vaccines. The preparation method of the tetravalent influenza virus split vaccine comprises the steps of chick embryo receiving and pre-incubation, chick embryo pre-irradiation inspection, virus inoculation and culture, chick embryo post-irradiation and embryo cooling, virus harvesting, ultrafiltration clarification, zone centrifugation and desugaring. Chromatography, virus splitting, virus inactivation, ultrafiltration liquid change, degerming filtration and preparation of a tetravalent influenza virus split vaccine finished product. Compared with the prior art, the technical means adopted by the invention effectively reduces the content of free formaldehyde and improves the vaccine antibody level.