INVESTIGADORES
SCHIJMAN Alejandro Gabriel
artículos
Título:
PCR-based screening and identification of Trypanosoma cruzi lineages in feces of triatomine species from rural northwestern Argentina.
Autor/es:
30 MARCET PL, DUFFY, T, CARDINAL MV, BURGOS JM, LAURICELLA MA, LEVIN MJ, KITRON U, GÜRTLER RE AND SCHIJMAN AG
Revista:
PARASITOLOGY
Referencias:
Año: 2006 vol. 132 p. 57 - 65
ISSN:
0031-1820
Resumen:
This study applied improved DNA extraction and polymerase chain reaction strategies for screening and identification of
Trypanosoma cruzi lineages directly from faeces of triatomines collected in a well-defined rural area in northwestern
Argentina. Amplification of the variable regions of the kinetoplastid minicircle genome (kDNA-PCR) was performed
in faecal lysates from 33 microscope (MO)-positive and 93 MO-negative Triatoma infestans, 2 MO-positive and 38
MO-negative Triatoma guasayana and 2 MO-positive and 73 MO-negative Triatoma garciabesi. kDNA-PCR detectedlineages directly from faeces of triatomines collected in a well-defined rural area in northwestern
Argentina. Amplification of the variable regions of the kinetoplastid minicircle genome (kDNA-PCR) was performed
in faecal lysates from 33 microscope (MO)-positive and 93 MO-negative Triatoma infestans, 2 MO-positive and 38
MO-negative Triatoma guasayana and 2 MO-positive and 73 MO-negative Triatoma garciabesi. kDNA-PCR detectedTriatoma infestans, 2 MO-positive and 38
MO-negative Triatoma guasayana and 2 MO-positive and 73 MO-negative Triatoma garciabesi. kDNA-PCR detectedTriatoma guasayana and 2 MO-positive and 73 MO-negative Triatoma garciabesi. kDNA-PCR detected
T. cruzi in 91% MO-positive and 7.5% MO-negative T. infestans, which were confirmed by amplification of the minicircle
conserved region. In contrast, kDNA-PCR was negative in all faecal samples from the other triatomine species. A panel of
PCR-based genomic markers (intergenic region of spliced-leader DNA, 24Sa and 18S rRNA genes and A10 sequence)
was implemented to identify the parasite lineages directly in DNA lysates from faeces and culture isolates from 28 infected
specimens. Two were found to be infected with TCI, 24 with TCIIe, 1 with TCIId and 1 revealed a mixed TCI+TCII
infection in the faecal sample whose corresponding culture only showed TCII, providing evidence of the advantages of
direct typing of biological samples. This study provides an upgrade in the current diagnosis and lineage identification ofin 91% MO-positive and 7.5% MO-negative T. infestans, which were confirmed by amplification of the minicircle
conserved region. In contrast, kDNA-PCR was negative in all faecal samples from the other triatomine species. A panel of
PCR-based genomic markers (intergenic region of spliced-leader DNA, 24Sa and 18S rRNA genes and A10 sequence)
was implemented to identify the parasite lineages directly in DNA lysates from faeces and culture isolates from 28 infected
specimens. Two were found to be infected with TCI, 24 with TCIIe, 1 with TCIId and 1 revealed a mixed TCI+TCII
infection in the faecal sample whose corresponding culture only showed TCII, providing evidence of the advantages of
direct typing of biological samples. This study provides an upgrade in the current diagnosis and lineage identification ofa and 18S rRNA genes and A10 sequence)
was implemented to identify the parasite lineages directly in DNA lysates from faeces and culture isolates from 28 infected
specimens. Two were found to be infected with TCI, 24 with TCIIe, 1 with TCIId and 1 revealed a mixed TCI+TCII
infection in the faecal sample whose corresponding culture only showed TCII, providing evidence of the advantages of
direct typing of biological samples. This study provides an upgrade in the current diagnosis and lineage identification of+TCII
infection in the faecal sample whose corresponding culture only showed TCII, providing evidence of the advantages of
direct typing of biological samples. This study provides an upgrade in the current diagnosis and lineage identification of
T. cruzi in field-collected triatomines and shows T. cruzi II strains as predominant in the region.in field-collected triatomines and shows T. cruzi II strains as predominant in the region.