Abstract

AGUILAR, Alicia et al. Use of the immunoperoxidase technique for the titration of a Dengue 1 virus strain. Vaccimonitor [online]. 2001, vol.10, n.1, pp.13-16. ISSN 1025-028X.

The use of the immunoperoxidase technique for the titration of Dengue virus has eliminated some of the disadvantages of plaque assay in cell culture for titration, which is considered a fastidious method due to the number of factors involved in the plaque formation. The time reduction to obtain results, is the principal advantage of the immunoperoxidase technique reported in the literature, but nothing is said about its precision. Our purpose was to test the immunoperoxidase technique to titrate Dengue 1 virus in our laboratory. The procedure described for this technique was followed, modified by the fixation of infected cells after 5, 7, and 10 days of incubation and by blocking with TSF-Tween 20- SBA 1% during one hour. The working dilution for the positive human sera was 1/500 and for the polyclonal ascitic fluid (AF) against Dengue 1 obtained in mice, 1/100. The useful dilutions for the anti human and anti mouse peroxidase conjugates were 1/100 and 1/200 for the first and 1/500 and 1/1000 for the second. In these conditions, it was possible to evidence the characteristic foci produced by Dengue 1 virus and to quantify it on the 7th and 10th days, but not on the 5th. The Variation Coefficient was < 30%. The 7th day was chosen as optimum to titrate the virus, leading us to titration faster than by the plaque method. Significant differences were not found between the viral titres obtained by both techniques.

Keywords : Dengue; immunoperoxidasa.

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