ELISA cualitativo de IgA anti-Lipopolisacárido de Vibrio cholerae en saliva de humanos.
Qualitative ELISA for IgA anti-Lipopolysaccharide of Vibrio cholerae in Human Saliva.
Judith Mónica del Campo, Rolando Ochoa, Miriam Lastre, Gustavo Bracho, Carlos Taboada, Miriam
Díaz y Oliver Pérez
Instituto Finlay. Centro de Investigación-Producción de Vacunas y Sueros. Ciudad de La Habana, Cuba. ]]>
E-mail: judithc@finlay.edu.cu
RESUMEN
Se estandarizó un ELISA para detectar el principal antígeno inductor de protección de Vibrio cholerae en saliva IgA contra el lipopolisacárido (LPS). El estudio se llevó a cabo en voluntarios que fueron inoculados por vía oral con dosis de 0, 107, 108, 109 unidades formadoras de colonias (ufc) del candidato vacunal El Tor Ogawa, cepa 638. Las muestras de saliva fueron tomadas de forma seriada, a los 0, 7, 8, 9, 10 y 14 días postinoculación. Se consideró seroconversión si las densidades ópticas eran superiores al nivel de corte y si los incrementos después de inmunizar duplicaban los valores antes de la inmunización. Los resultados, al ser comparados con los grupos experimentales, condición individuos inoculados y los placebos, demostraron que la técnica tiene una sensibilidad del 93,3%, una especificidad del 96,0%, un valor predictivo positivo de 98,2% y negativo de 85,7%, y una eficiencia del 94,1%. Se demostró la presencia de IgA anti LPS en saliva de los individuos inoculados con el candidato vacunal, con una mayor concentración de anticuerpos con el inóculo de (109 ufc) y se obtuvo la máxima positividad a los nueve días.
Palabras claves: Cólera, saliva, mucosa, IgA, ELISA
ABSTRACT
An ELISA technique for IgA antibodies again V. cholerae lipopolisaccharide (LPS) in saliva was standardized. Humans volunteers were orally immunized with 0, 107, 108, 109 colony forming units of vaccine candidate 638, El Tor Ogawa. Saliva samples were collected 0, 7, 8, 9, 10 and 14 days after oral administration.A Seroconversion was considered, if IgA titers determinated by optical density, were higher than the cutoff value, and if increase after immunization duplicate the values before immunization. The technique showed a sensitivity of 93.3%, a specificity of 96.0% a positive predictive value of 98.2%, a negative predictive value of 85.7% and an efficiency of 94.1% when they were compared to the experimental group. The presence of specific IgA against LPS in saliva was demonstrated after immunization with the vaccine candidate. The results showed a kinetic profile with a maximum of IgA titter at day 9 post immunization with the highest frequency of positive titers found in the group immunized with 109 cfu.
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