Examinando por Autor "Duque Varela, Juan Pablo"
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Ítem Estandarización de una metodología para la cuantificación de títulos virales por citometría de flujo: virus dengue y zika(Universidad El Bosque) Almanza Holguín, Laura Natalia; Duque Varela, Juan Pablo; Delgado Tiria, Felix Giovanni; Delgado Tiria, Felix Giovanni [0000-0001-6685-7507]The basic research in virology has a Standard Gold methodology for the calculation of viral titer which is the plating technique. However, this technique requires a wide availability of time and experiences by the experimenter; Therefore, proposing an alternative and equally reliable technique is a challenge that currently exists. For this reason, this project aims to statistically compare the results obtained between the traditional method and flow cytometry to see if it is feasible to use the latter as a routine technique in the quantification of viral loads. For this, three different cell lines (VERO, BHK-21 and U937-DC-SIGN) and the dengue (4 serotypes) and Zika viruses were used in a new protocol for flow cytometry which is based on the use of 2-fold serial dilutions of viral inoculum in order to obtain percentages of infected cells and then calculate the viral titers. The results obtained by flow cytometry were compared with those previously obtained by plaque assay, applying a correlation analysis. As a result, it was evident that the viral titers calculated using the BHK-21 cells correlated significantly with those reported by the plaque assay. Additionally, it was observed that the infectivity curves were different between the different cell lines used, showing different degrees of susceptibility to infection at the time of the test. In conclusion, our results show that the flow cytometry technique can be used as an alternative method for the quantification of viral particles, obtaining results that correlate with those obtained by plating, as long as BHK-21 cells are used.