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The cultured Enzyme-Linked ImmunoSpot (ELISpot) assay is a functional T cell assay, which is commonly used to assess virus-specific T cell responses. The use of an in vitro expansion step before the ELISpot distinguishes such "cultured" ELISpots from "ex vivo" ELISpots. Cultured ELISpots have the advantage that lower frequency responses can be analyzed compared to ex vivo ELISpots, but do carry the associated potential distortions of the expansion phase. Cultured ELISpot assays are of value to determine silent and symptomatic transmission of the Dengue virus (DENV) in the community and to identify the correlates of a DENV-specific protective immune response. We have evaluated T cell responses to the DENV using cultured ELISpot assays with serotype-specific T cell epitopes to determine past infecting dengue virus (DENV) serotypes. The peptides used in this assay do not cross react with the Japanese encephalitis virus nor other flaviviruses. Therefore, this assay is likely to be useful in determining the past infecting DENV serotype in immune-epidemiological studies and in dengue vaccine trials.

Original publication




Journal article


Methods Mol Biol

Publication Date





165 - 171


Cultured ELISpot assay, Dengue virus, Serotype-specific T cell responses, Short-term T cell cultures, Cells, Cultured, Dengue, Dengue Virus, Enzyme-Linked Immunospot Assay, Epitopes, T-Lymphocyte, Humans, Leukocytes, Mononuclear, Peptides, T-Lymphocytes