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Development and assessment of an improved recombinant multiepitope antigen-based immunoassay to diagnose chronic Chagas disease

Published online by Cambridge University Press:  28 March 2018

Luz María Peverengo
Affiliation:
Laboratorio de Tecnología Inmunológica (Facultad de Bioquímica y Ciencias Biológicas-Universidad Nacional del Litoral), Santa Fe, Argentina
Valeria Garcia
Affiliation:
INTEC (CONICET-UNL), Santa Fe, Argentina
Luz María Rodeles
Affiliation:
Laboratorio de Tecnología Inmunológica (Facultad de Bioquímica y Ciencias Biológicas-Universidad Nacional del Litoral), Santa Fe, Argentina Laboratorio de Investigación en Ciencias Biomédicas (Facultad de Ciencias Médicas–Universidad Nacional del Litoral), Santa Fe, Argentina
Diego Mendicino
Affiliation:
Centro de Investigaciones sobre Endemias Nacionales (FBCB-UNL), Santa Fe, Argentina
Miguel Vicco
Affiliation:
Laboratorio de Investigación en Ciencias Biomédicas (Facultad de Ciencias Médicas–Universidad Nacional del Litoral), Santa Fe, Argentina
Claudia Lagier
Affiliation:
Dpto. de Quimica Analitica (FBIOyF-UNR), Rosario, Argentina
Verónica Gonzalez
Affiliation:
INTEC (CONICET-UNL), Santa Fe, Argentina
Luis Gugliotta
Affiliation:
INTEC (CONICET-UNL), Santa Fe, Argentina
Iván Marcipar*
Affiliation:
Laboratorio de Tecnología Inmunológica (Facultad de Bioquímica y Ciencias Biológicas-Universidad Nacional del Litoral), Santa Fe, Argentina Laboratorio de Investigación en Ciencias Biomédicas (Facultad de Ciencias Médicas–Universidad Nacional del Litoral), Santa Fe, Argentina
*
Author for correspondence: Luz María Peverengo, E-mail: luzpeverengo@gmail.com

Abstract

The use of chimeric molecules fusing several antigenic determinants is a promising strategy for the development of low-cost, standardized and reliable kits to determine specific antibodies. In this study, we designed and assessed a novel recombinant chimera that complements the performance of our previously developed chimera, CP1 [FRA and SAPA antigens (Ags)], to diagnose chronic Chagas disease. The new chimeric protein, named CP3, is composed of MAP, TcD and TSSAII/V/VI antigenic determinants. We compared the performance of both chimeric Ags using a panel of 67 Trypanosoma cruzi-reactive sera and 67 non-reactive ones. The sensitivity of CP3 vs CP1 was 100 and 90.2%, and specificity was 92.5 and 100%, respectively. The mixture of CP1 + CP3 achieved 100% of sensitivity and specificity. More importantly, an additional subset of 17 sera from patients with discordant results of conventional serological methods was analysed; the CP1 + CP3 mixture allowed us to accurately classify 14 of them with respect to IIF, the usual technique used in most of the reference centres. These results show an improved performance of the CP1 + CP3 mixture in comparison with enzyme-linked immunosorbent assay and indirect haemagglutination commercial assays.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2018 

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