Human rotavirus-A seed strain ZTR-68, and isolation, culture and identification thereof
A rotavirus and strain technology, applied in the directions of virus/phage, analytical materials, inactivation/attenuation, etc., can solve the problems of high cost of cold chain storage system, intussusception and virulence reversion mutation, etc. The effect of improving the efficiency of primary culture, high virus recovery yield, and high impurity removal effect
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[0033] Embodiment 1, the separation method of ZTR-68 strain involved in the present invention:
[0034] (1) Obtain virus samples
[0035] Collect diarrhea excrement samples from pediatric diarrhea patients in the hospital, resuspend in 0.01 M PBS, pH 7.2 solution, centrifuge at 12000 g for 20 minutes to get the supernatant, and use a 0.22 μm filter (purchased from Millipore) to sterilize and filter,- Store at 20°C for later use.
[0036] (2) Adaptation of virus on MA104 cells
[0037] Use 0.25% trypsin-EDTA to digest the P 29 generation MA104 cells that have formed a monolayer. 5.0 cells / ml. Press 2×10 5.0 Cells / bottle were inoculated into a small square bottle, and after culturing at 37°C for 3 to 4 days, inoculated 200 μl of the diluent of diarrhea samples from rotavirus-infected children on the cell monolayer, adsorbed at 37°C for 2 hours, and then added calf serum-free DMEM cell maintenance solution, cultivated at 37°C and observed cell changes. After 3 to 4 days of cy...
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