PCR-based screening and lineage identification of Trypanosoma cruzi directly from faecal samples of triatomine bugs from northwestern Argentina
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 kineto...
Guardado en:
| Publicado: |
2006
|
|---|---|
| Materias: | |
| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00311820_v132_n1_p57_Marcet http://hdl.handle.net/20.500.12110/paper_00311820_v132_n1_p57_Marcet |
| Aporte de: |
| id |
paper:paper_00311820_v132_n1_p57_Marcet |
|---|---|
| record_format |
dspace |
| spelling |
paper:paper_00311820_v132_n1_p57_Marcet2023-06-08T14:56:55Z PCR-based screening and lineage identification of Trypanosoma cruzi directly from faecal samples of triatomine bugs from northwestern Argentina Chagas disease Lineage PCR Triatoma garciabesi Triatoma guasayana Triatoma infestans Trypanosoma cruzi Argentina article cell culture cell isolation cell lineage Chagas disease feces analysis genome human marker gene nonhuman parasite parasite identification polymerase chain reaction priority journal protozoal infection sample Triatoma Trypanosoma cruzi Kinetoplastida Triatoma Triatoma garciabesi Triatoma guasayana Triatoma infestans Trypanosoma cruzi 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 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, 24Sα 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 T. cruzi in field-collected triatomines and shows T. cruzi II strains as predominant in the region. © 2005 Cambridge University Press. 2006 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00311820_v132_n1_p57_Marcet http://hdl.handle.net/20.500.12110/paper_00311820_v132_n1_p57_Marcet |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Chagas disease Lineage PCR Triatoma garciabesi Triatoma guasayana Triatoma infestans Trypanosoma cruzi Argentina article cell culture cell isolation cell lineage Chagas disease feces analysis genome human marker gene nonhuman parasite parasite identification polymerase chain reaction priority journal protozoal infection sample Triatoma Trypanosoma cruzi Kinetoplastida Triatoma Triatoma garciabesi Triatoma guasayana Triatoma infestans Trypanosoma cruzi |
| spellingShingle |
Chagas disease Lineage PCR Triatoma garciabesi Triatoma guasayana Triatoma infestans Trypanosoma cruzi Argentina article cell culture cell isolation cell lineage Chagas disease feces analysis genome human marker gene nonhuman parasite parasite identification polymerase chain reaction priority journal protozoal infection sample Triatoma Trypanosoma cruzi Kinetoplastida Triatoma Triatoma garciabesi Triatoma guasayana Triatoma infestans Trypanosoma cruzi PCR-based screening and lineage identification of Trypanosoma cruzi directly from faecal samples of triatomine bugs from northwestern Argentina |
| topic_facet |
Chagas disease Lineage PCR Triatoma garciabesi Triatoma guasayana Triatoma infestans Trypanosoma cruzi Argentina article cell culture cell isolation cell lineage Chagas disease feces analysis genome human marker gene nonhuman parasite parasite identification polymerase chain reaction priority journal protozoal infection sample Triatoma Trypanosoma cruzi Kinetoplastida Triatoma Triatoma garciabesi Triatoma guasayana Triatoma infestans Trypanosoma cruzi |
| description |
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 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, 24Sα 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 T. cruzi in field-collected triatomines and shows T. cruzi II strains as predominant in the region. © 2005 Cambridge University Press. |
| title |
PCR-based screening and lineage identification of Trypanosoma cruzi directly from faecal samples of triatomine bugs from northwestern Argentina |
| title_short |
PCR-based screening and lineage identification of Trypanosoma cruzi directly from faecal samples of triatomine bugs from northwestern Argentina |
| title_full |
PCR-based screening and lineage identification of Trypanosoma cruzi directly from faecal samples of triatomine bugs from northwestern Argentina |
| title_fullStr |
PCR-based screening and lineage identification of Trypanosoma cruzi directly from faecal samples of triatomine bugs from northwestern Argentina |
| title_full_unstemmed |
PCR-based screening and lineage identification of Trypanosoma cruzi directly from faecal samples of triatomine bugs from northwestern Argentina |
| title_sort |
pcr-based screening and lineage identification of trypanosoma cruzi directly from faecal samples of triatomine bugs from northwestern argentina |
| publishDate |
2006 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00311820_v132_n1_p57_Marcet http://hdl.handle.net/20.500.12110/paper_00311820_v132_n1_p57_Marcet |
| _version_ |
1768542352976117760 |