I have isolated dengue virus from infected patient serum and for titration i did plaque assay by using different conc. of vero cells in 12well as well as in 24 well plate, but i didnt get plaques.
if someone suggest me a proper protocol, that would be very helpul for me to proceed further..thanks..
Quantification of DENV titers by lytic plaque assays is commonly performed using Vero or BHK21 cells. The procedure is basically the same. It is often used SeaKem ME agarose. Particularly in our laboratory we use molecular biology agarose with satisfactory results. You may consult these references: Diamond et al. J Virol. 2000 Sep;74(17):7814-23 and Medina et al. Curr Protoc Microbiol. 2012 Nov;Chapter 15:Unit 15D.2.
This review article will help you
Guidelines for Plaque-Reduction Neutralization Testing of Human Antibodies to Dengue Viruses.
Roehrig JT, Hombach J, Barrett AD.
Viral Immunol. 2008 Jun;21(2):123-32. doi: 10.1089/vim.2008.0007. Review.
PMID: 18476771
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