Revista Peruana de Medicina Experimental y Salud Publica
versión impresa ISSN 1726-4634
YABAR V, Carlos; CHOQUE P, Juana y MONTOYA P, Ysabel. Evaluación serológica de una proteína recombinante a partir de una cepa aislada del virus de la fiebre amarilla en el Perú: un estudio piloto. Rev. perú. med. exp. salud publica [online]. 2003, vol.20, n.4, pp. 193-199. ISSN 1726-4634.
Objective: To assess a 66kDa (Er66) recombinant protein of yellow fever (YF) virus using immunoreactive antibodies. Material and methods: Er66 was expressed in Escherichia coli and then it was challenged against 1/100, 1/50 and 1/25 monoclonal antibodies containing dilutions, and against clinical sera containing specific IgM and IgG antibodies against YF virus using western blot assay. Likewise, total protein content from yellow fever virus and Dengue (DEN) virus were also tested as antigenicity control. Results: Er66 recombinant protein showed antigenicity against 1/50 and 1/25 titers of monoclonal antibodies (MAB8701); however, no immunoreactivity was observed in sera positive for yellow fever and dengue infections. Serological assays with crude extracts of 17D Asibi yellow fever virus strain revealed that antigenicity of 60 - 80 kDa viral proteins was abrogated at 100 °C (denaturizing temperature). In contrast, when total protein was subjected to b-mercaptoethanol treatments without heat, the antigenicity increased. Using this principle, ER66 protein was reduced under b-mercaptoethanol treatment and it was newly challenged to reactive sera. The final result revealed that Er66 generated immunoreactivity in sera with high titers of IgM and IgG antibodies for yellow fever, while other samples showed a weak immunoreactivity signal. Conclusion: This study shows that Er66 was recognized by specific IgM and IgG yellow fever antibodies from clinical samples of infected patients. The data presented in this study suggest the use of recombinant proteins as possible candidates with a diagnostic value for yellow fever.
Palabras llave : Yellow fever virus; Serology; Recombinant Proteins; Monoclonal Antibodies; Peru.