ESTABLISHMENT OF PLAQUE ASSAY METHOD FOR TITRATING YELLOW FEVER VACCINE
- Central Research Institute, Kasauli.
- Central Drugs Laboratory, Kasauli.
- Bhojia Institute, Baddi.
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The present study was undertaken with a view to look for the use of Vero cell line for plaque assay of Yellow fever vaccine, over Mouse LD50 method, for vaccine potency test. To establish the plaque assay the growth conditions for Vero cell line were standardized. Different parameters like cell count, dilution, inoculum size, adsorption time for virus, concentration for Methyl Cellulose (MC) in the overlay media and incubation period were standardized. And titration of Yellow fever vaccine was performed by Plaque Forming Unit (PFU) method. A cell concentration of 3-4?105 cells per ml, in tissue culture plate gave a complete monolayer after overnight incubation. Virus dilution of 10-2 was optimum for desired pfu/0.5ml. An inoculum size of 0.2 ml per well was found suitable. Maximum plaques were obtained after an adsorption time of 4 hours, at 36 ?1?C, by 7 days of incubation. Different concentrations of methyl cellulose, overlay media were used. 1% methyl cellulose in Minimum Essential Media (MEM) with 2% FBS gave good results. The different samples of Yellow fever 17D vaccines were titrated by Plaque Assay method along with the IRS and titre results were compared with the titre values of manufacturer?s potency assay. The vaccine pass the limit of minimum requirement of potency i.e., 4.24 log10 PFU/0.5ml dose, equivalent to titre 1000 MouseLD50.
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[Ankita Semwal, Pallavi Kapoor, Kanika Sharma, Shweta Tandon and Swati Chaudhary. (2017); ESTABLISHMENT OF PLAQUE ASSAY METHOD FOR TITRATING YELLOW FEVER VACCINE Int. J. of Adv. Res. 5 (Mar). 2336-2346] (ISSN 2320-5407). www.journalijar.com
Central Research Institute, Kasauli.
