The chicken embryo culture step of the virus.

First, the purpose requires mastering the operation technology of the chicken embryo urinary sac cavity vaccination to cultivate the Newtown epidemic virus.
, equipment 1. Chicken embryo should choose a healthy flock of fertilized eggs, inoculated chicken embryo should be no henogeneic antibodies, so as not to affect the proliferation of the virus in the embryo.
used in experiments with chicken embryos are mostly 9-10 days old live embryos.
2. Vaccination material Newtown disease I vaccine 3. Various instruments Incubators, egg boxes, egg racks, punchers, 1 ml TB mycosin syringes, 41/2 injection needles, tweezers, 3% iodine tarts, 75% alcohol cotton balls, pencils, alcohol lamps, scissors, eye tweezers, sterilized vaccine bottles, sterilized drop tubes and solid paraffins.
3. Content and methods (i) pre-vaccination preparation 1. Preparation of chicken embryo This experiment selects 9-10 days old chicken embryo, in the egg with a pencil to draw out the gas chamber, in the gas chamber slightly below the embryo active and blood vessels obvious area, in the gap between blood vessels marked No., as the vaccination site; wash the eggshell clean and dry with 0, 1% new chelation, and then disinfect the inoculation site with 2% iodine, wipe it again with 75% alcohol, punch the hole in the inoculation positioning, and place the chamber up on the egg rack.
2. The preparation of the virus inoculation material to isolate the cultured viral material should be sterile, so in order to achieve the sterile purpose of the material, penicillin and streptomycin can be added to each ml of inoculation substances 100-500IU, 1 hour at room temperature or 12-24 hours in the refrigerator treatment.
or filter sterilization.
(ii) Inoculation fluffy urine sac cavity injection: 1 ml syringe with the 6th needle to absorb the Newtown disease virus material, needle from the well-drilled hole into the direction of the chicken embryo 0.5--1cm, injection 0, 1 ml (quite 0, 5 plumes), then add zinc oxide cloth seal, then add melted solid paraffin seal, put back in the incubator for hatching, double eggs per day, take care of a .24 hours of death.
(iii) harvest new city disease I virus materials, in the vaccination 24-48 hours can be harvested.
the chicken embryo in a 4-degree refrigerator for 4 hours or overnight before harvesting to avoid ruptured blood vessels during anatomy.
iodine wine disinfection eggshell, with tweezers to the eggshell and urine sac membrane, with a suction head to suck urine sac in the bottle, can collect 5-8 ml, sterile examination, frozen preservation.
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