Fluorometric cell-ELISA for quantifying rabies infection and heparin inhibition
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Fecha
2005
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Publicado en
Journal of virological methods, 1879-0984, Vol. 127, Nro. 1, 2005, p. 33-39
Publicado por
Elsevier
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Resumen
Descripción
Abstract
The purpose of the present study was to implement a fluorometric method for detecting and quantifying viral antigens in human meduloblas-toma cells infected by two types of fixed rabies virus (CVS-MB and CVS-BHK) and a street virus using a cell-enzyme linked immunosorbentassay (cell-ELISA) technique; alkaline phosphatase was used as the antibody-marker enzyme and 4-methyl-umbelliferyl-phosphate as thefluorogenic substrate. The system was used for detecting up to 1:10,000 viral inoculums, followed by evaluating the effect of heparin oninfection. Infected cultures were reliably differentiated from their respective negative controls in both assays allowing data to be analysedstatistically. As reported in another study, heparin produces strong inhibition when the CVS-BHK viral strain is used for infection; it has thusbeen suggested that it binds to the neural cell adhesion molecule and could be blocked by using this drug. This fluorometric method is lesstime-consuming, has increased reproducibility and useful for quantitation of collected data and can therefore be considered as a useful toolfor research.
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Keywords
4-Methyl-umbelliferyl-phosphate (MUP), Cell-ELISA, Fluorometric method, Heparin, Human meduloblastoma cells, Rabies virus