Propagation and Validation of Dengue and Chikungunya Viruses

We used the clinical isolates dengue ST (passage 6) [35 (link)] and Chikungunya EAS DMERI09/08 (passage 3) viruses, collected from the Singapore General Hospital in 1997 and National University Hospital, Singapore in 2008, respectively (already existing collection). A portion of the E1 gene from CHIKV EAS was amplified and sequenced to validate the absence of the A226V amino acid substitution [9 (link)]. All viruses were propagated in Vero cells (ATCC) and titrated by plaque assay in BHK cells (ATCC) as previously described [36 (link)].

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Mendenhall I.H., Manuel M., Moorthy M., Lee T.T., Low D.H., Missé D., Gubler D.J., Ellis B.R., Ooi E.E, & Pompon J. (2017). Peridomestic Aedes malayensis and Aedes albopictus are capable vectors of arboviruses in cities. PLoS Neglected Tropical Diseases, 11(6), e0005667.

Publication 2017

Vero cells BHK cells

A gene Amino acid substitution Assay Cells Chikungunya Dengue Plaque Vero cells Viruses

Corresponding Organization : Centre National de la Recherche Scientifique

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Variable analysis

independent variables

Dengue ST (passage 6) virus
Chikungunya EAS DMERI09/08 (passage 3) virus

dependent variables

Viral propagation in Vero cells
Viral titration by plaque assay in BHK cells

control variables

Vero cells (ATCC)
BHK cells (ATCC)

controls

Absence of the A226V amino acid substitution in the E1 gene of CHIKV EAS was validated

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