Medicine, disinfectant and additive for resisting porcine pseudorabies virus and using method of medicine, disinfectant and additive
A technology for porcine pseudorabies and disinfectant, which is applied in the field of disinfectants and drugs against porcine pseudorabies virus, and can solve the problems of affecting the entry of the virus, affecting the replication of the virus, and enhancing the virulence of the mutant strain to the pig herd.
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Embodiment 1
[0030] PK-15 cells recovered from our laboratory were kept in DMEM containing 5% fetal bovine serum at 37°C 5% CO 2 Cultured in an incubator, and passed to the third generation, with 5×10 per well 5 Evenly spread the concentration of each cell in a 6-well plate. After culturing to the logarithmic growth phase, after about 16 hours, when the cells grow to about 70% to 75% of the volume of each well, discard the DMEM containing 5% fetal bovine serum, PBS Wash the buffer three times, absorb the remaining PBS in the last pass, add 1ml of DMEM and dilute the dissolved Huaier polysaccharide to 25mg, 50mg, 100mg, 200mg, incubate PRV XJ5 (MOI=0.1) with the cells for 1h, and change after 1h Prepare 2ml of DMEM solution containing 2% fetal bovine serum, and add PBS buffer or corresponding concentration of Huaier polysaccharide (25mg, 50mg, 100mg, 200mg), and observe the cell morphology and pathological changes under the microscope at 12h and 24h respectively. according to figure 1 The...
Embodiment 2
[0032] (1) Western blot detects the activity of Huaier polysaccharide on PK-15 cells in inhibiting the infection of porcine pseudorabies virus mutant strain XJ5 strain:
[0033]PK-15 cells were digested with trypsin and diluted with DMEM nutrient solution with a volume concentration of 5% fetal bovine serum to 5×10 5 The concentration per well was added dropwise to a 6-well plate and placed at 37°C, 5% CO 2 After the cells adhered to the wall and formed a monolayer in the logarithmic growth phase in the incubator (about 17h), wash the cells three times with PBS buffer, and after absorbing the residual PBS, add Sophora japonica diluted to the corresponding concentration with 1mL serum-free DMEM. Ear polysaccharides (25μg, 50μg, 100μg, 200μg) and porcine pseudorabies virus variant XJ5 strain were mixed with PK-15 cells at 37°C, 5%CO 2 After incubating in the incubator for 1 hour, replace it with 2 mL of DMEM nutrient solution containing 2% fetal bovine serum and maintain the co...
Embodiment 3
[0039] (1) Western blot detection of Huaier polysaccharides on PK-15 cells to inhibit the adsorption and entry of porcine pseudorabies virus variant XJ5 strain:
[0040] PK-15 cells were digested with trypsin and diluted with DMEM nutrient solution with a volume concentration of 5% fetal bovine serum to 5×10 5 The concentration per well was added dropwise to a 6-well plate and placed at 37°C, 5% CO 2 After the cells adhered to the wall and formed a monolayer in the logarithmic growth phase in the incubator (about 17h), the cells were washed three times with 4°C PBS buffer, and after the residual PBS was exhausted, add 1 mL of serum-free DMEM at 4°C to dilute to Corresponding concentrations of Huaier polysaccharide (25 μg, 50 μg, 100 μg, 200 μg) and porcine pseudorabies virus mutant strain XJ5 were incubated with PK-15 cells at 4°C for 1 h, and then replaced with 2 mL of DMEM nutrient solution containing 2% fetal bovine serum. at 37°C, 5% CO 2 Incubate in an incubator for 1 h...
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