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Physiological, morphological, and immunochemical parameters used for the characterization of clinical and environmental isolates of Acanthamoeba

Published online by Cambridge University Press:  09 November 2012

A. BECKER-FINCO ,
A. O. COSTA ,
S. K. SILVA ,
J. S. RAMADA ,
C. FURST ,
A. E. STINGHEN ,
B. C. DE FIGUEIREDO ,
J. DE MOURA  and
L. M. ALVARENGA
A. BECKER-FINCO
Affiliation:
Departamento de Patologia Básica, UFPR, Curitiba, Paraná, Brazil
A. O. COSTA
Affiliation:
Departamento de Análises Clínicas e Toxicológicas, Faculdade de Farmácia, UFMG, Belo Horizonte, Minas Gerais, Brazil
S. K. SILVA
Affiliation:
Departamento de Patologia Básica, UFPR, Curitiba, Paraná, Brazil
J. S. RAMADA
Affiliation:
Departamento de Patologia Básica, UFPR, Curitiba, Paraná, Brazil
C. FURST
Affiliation:
Departamento de Patologia, Centro de Ciências da Saúde, UFES, Vitória, Espírito Santo, Brazil
A. E. STINGHEN
Affiliation:
Departamento de Patologia Básica, UFPR, Curitiba, Paraná, Brazil
B. C. DE FIGUEIREDO
Affiliation:
Instituto Pelé Pequeno Príncipe, Faculdades Pequeno Príncipe, Curitiba, Paraná, Brazil
J. DE MOURA
Affiliation:
Departamento de Patologia Básica, UFPR, Curitiba, Paraná, Brazil
L. M. ALVARENGA*
Affiliation:
Departamento de Patologia Básica, UFPR, Curitiba, Paraná, Brazil
*
*Corresponding author: Departamento de Patologia Básica, UFPR, Curitiba, Paraná, Brazil. Tel: +55 41 3361 1692. Fax: +55 41 3266 2042. E-mail: [email protected]
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Summary

The factors that characterize Acanthamoeba strains as harmless or potentially pathogenic have not been elucidated. Analysing the in vitro and in vivo parameters of Acanthamoeba samples, including heat tolerance at temperatures close to that of the human body, cytopathic effects, and their ability to cause infections in animals, has been proposed to identify their pathogenic potential. Another promising criterion for differentiating strains is the analysis of their biochemical and immunochemical properties. In this study, a comparative evaluation between clinical and environmental Acanthamoeba isolates was performed on the basis of physiological, morphological, and immunochemical criteria. Crude antigens were used to characterize the protein profiles by electrophoresis and immunize mice to produce polyclonal and monoclonal antibodies. The antibodies were characterized using ELISA, Western blotting, and immunofluorescence techniques. The results obtained with polyclonal antibodies suggest the presence of specific proteins for each studied isolate and co-reactive immunochemical profiles among conserved components. Ten monoclonal antibody clones were obtained; mAb3 recognized 3 out of 4 samples studied. The results of this study may help standardize criteria for identifying and characterizing Acanthamoeba strains. Taken together, our results support the view that a set of features may help differentiate Acanthamoeba species and isolates.

Keywords

Acanthamoebaamoebic keratitismonoclonal antibodiespolyclonal antibodiesprotein profileenvironmental strain
Type
Research Article
Information
Parasitology , Volume 140 , Issue 3 , March 2013 , pp. 396 - 405
Copyright
Copyright © Cambridge University Press 2012

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