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Vaccimonitor

On-line version ISSN 1025-0298

Abstract

PEREZ-BARRIOS, Yosbany et al. Standardization of an ELISA for the quantification of IgG antibodies against outer membrane vesicle of Salmonella Paratyphi A. Vaccimonitor [online]. 2018, vol.27, n.3, pp. 76-83. ISSN 1025-0298.

Salmonella Paratyphi A, is an exclusive pathogen of humans, being the second most common cause of enteric fever in Southeast Asia. Recently the incidence of this disease in this continent has increased, displacing Salmonella enterica serotype Typhi as the first cause of enteric fever. Currently there are no vaccines licensed against S. Paratyphi A. The Finlay Institute of Vaccines is working on obtaining a vaccine candidate based on outer membrane vesicles (VME) against S. Paratyphi A, so it became necessary to develop a technique for the evaluation of its immunogenicity. The objective of this work was the standardization of an ELISA for the quantification of IgG antibodies against VME of S. Paratyphi A. The best conditions of this assay were determined in terms of optimum concentration of coating and working dilution of the conjugate. In addition, the interval and linearity of the curve, the intra- and inter-assay precision, the specificity and the limit of detection were defined. The calibration curve was generated with an internal standard serum and presented a good linear fit with an R2 =0.98. The coefficients of variation in the intra- and interassay precision tests were in the intervals established for each one (=10%, =20% respectively). The results obtained support the use of this quantitative ELISA for the evaluation of the immunogenicity of VME formulations of S. Paratyphi A in research and development phases

Keywords : ELISA; standardization; Salmonella Paratyphi A; Vaccines; Outer Membrane Vesicles.

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