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Normalización de la técnica de neutralización por placas en las células Vero para los virus del dengue

INTRODUCTION: the standard plaque reduction neutralization technique has been performed in BHK-21 cells for more than 20 years in Cuba to determine the neutralizing antibodies to dengue. At the end of 2007, the WHO implemented a program to harmonize this technique at all the laboratories worldwide. OBJECTIVES: the present study was aimed at standardizing the plaquereduction neutralization technique in Vero cells from the Cultural Lab of “Pedro Kouri” Tropical Medicine Institute. METHODS: viral strains suggested by Who, strains from the Strain Bank of Arbovirus Laboratory, as well as the panel of human sera from dengue-suspected patients after 5 days of the symptoms. RESULTS: it was proved that suspensioncultured Vero cells allow higher plaque capacity and quality. Similar results were reached using Arbovirus Bank strains and WHO reference strains. CONCLUSIONS: it was possible to standardize the plaque-reduction neutralization in Vero cells, with results similar to those of BHK-21 but in longer length of time. Standardization of this technique will allow comparing the outcome of neutralization in the epidemiological studies and vaccine research with those of other laboratories worldwide.