Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/14679
Title: Trypanosoma cruzi: parasite detection and strain classification in chronic chagasic patients from North-Eastern Brazil using PCR amplification of kinetoplast DNA and non-radioactive hybridization
Authors: Britto, C.
Cardoso, M. A.
Ravel, C.
Santoro, A.
Pereira, J. Borges
Coura, J. R.
Morel, Carlos Medicis
Winckler, P.
Affilliation: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bioquímica e Biologia Molecular. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bioquímica e Biologia Molecular. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.
Université de Montpellier. Faculte de Medicine. Laboratoire de Parasitologie. Montpellier, France.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Medicina Tropical. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Medicina Tropical. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Medicina Tropical. Rio de Janeiro, RJ, Brasil
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bioquímica e Biologia Molecular. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bioquímica e Biologia Molecular. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.
Abstract: Blood samples from 172 individuals from northeastern Brazil were subjected to PCR amplification of Trypanosoma cruzi-specific kDNA sequences. This method enabled us to detect parasite DNA in 21 of 47 patients that were serologically positive, In addition, 1 patient that gave doubtful results with chagasic serology was confirmed as positive by PCR. We applied the same PCR detection method to the feces of wild triatomines captured in the same region, obtaining three positive results that were confirmed by microscopic examination. The 25 amplified products obtained in this study were then reamplified with primers that gave a final amplicon containing sequences from the most variable region of kDNA minicircles. These were used as probes in hybridization experiments aimed at defining the degree of relatedness between the strains infecting humans and insects based on kDNA homologies. We found that the amplification products from the three triatomines were related and showed no crosshybridization with those obtained from human infections. Eight amplified products from human infections showed no cross-hybridization and did not hybridize with products from other patients. This indicates that the strains of T. cruzi circulating in the region present a high level of genetic heterogeneity. Finally, a number of amplified products hybridized with amplicons that did not hybridize with each other, indicating that infections with a parasite population presenting a mixed kDNA content (either due to different strains of T. cruzi or to a hybrid parasite) are a more frequent event than previously thought.
Keywords: Trypanosoma cruzi
Chronic Chagasic Patients
Northeastern Brazil
PCR diagnosis
Kinetoplast DNA
Nonradiactive hybridization
keywords: Nordeste do Brasil
DeCS: Trypanosoma cruzi
Doença de Chagas
DNA de Cinetoplasto
Reação em Cadeia da Polimerase
Issue Date: 1995
Publisher: Elsevier
Citation: BRITTO, C. et al. Trypanosoma cruzi: parasite detection and strain classification in chronic chagasic patients from North-Eastern Brazil using PCR amplification of kinetoplast DNA and non-radioactive hybridization. Experimental Parasitology, v. 81, p. 462-471, 1995.
ISSN: 0014-4894
10.1006/expr.1995.1139
Copyright: open access
Appears in Collections:IOC - Artigos de Periódicos

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