Detection of Histoplasma Capsulatum by Nested-PCR in Clinical Specimens
Histoplasmosis is a systemic mycosis whose diagnosis is based on clinical, mycological and immunological criteria. Identification of its etiological agent in biological samples is difficult to perform. We evaluated a nested-PCR technique to identify Histoplasma capsulatum in clinical specimens (bloo...
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Universidad del Zulia,Facultad de Medicina,Departamento de Enfermedades Infecciosas y Tropicales
2007
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ftdoajarticles:oai:doaj.org/article:0d31ff9116a94e549776ef6d2b4a1900 2023-10-01T03:54:07+02:00 Detection of Histoplasma Capsulatum by Nested-PCR in Clinical Specimens María Teresa Colella Sofía Mata Claudia Hartung Celina Pérez Arantza Roselló Carolina Olaizola Carmen Fernández Silvia Magaldi Félix Toro 2007-12-01T00:00:00Z https://doaj.org/article/0d31ff9116a94e549776ef6d2b4a1900 EN ES eng spa Universidad del Zulia,Facultad de Medicina,Departamento de Enfermedades Infecciosas y Tropicales https://produccioncientificaluz.org/index.php/kasmera/article/view/4794 https://doaj.org/toc/0075-5222 https://doaj.org/toc/2477-9628 0075-5222 2477-9628 https://doaj.org/article/0d31ff9116a94e549776ef6d2b4a1900 Kasmera, Vol 35, Iss 2, Pp 156-163 (2007) histoplasmosis PCR ADN Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2007 ftdoajarticles 2023-09-03T00:41:48Z Histoplasmosis is a systemic mycosis whose diagnosis is based on clinical, mycological and immunological criteria. Identification of its etiological agent in biological samples is difficult to perform. We evaluated a nested-PCR technique to identify Histoplasma capsulatum in clinical specimens (blood and bone marrow) and reference strains. Three different methods for DNA isolation were tested based on the reagents Guanidine thiocyanate, Benzoyl chloride and Triton X-100. Among these procedures, Guanidine thiocyanate-based method yielded by utmost good qualityDNAand was easy to perform when used with clinical specimens. However, a better recovery of DNA was accomplished with the Triton-X-100 method. Reference strains of Paracoccidioides brasiliensis, Candida glabrata, Cryptococcus neoformans and Coccidioides immitis (C posadasii) were tested to assess PCR assay specificity. The samples were amplified by nested-PCR and analyzed by 3% agarose gel electrophoresis. Each of the reference strains showed the same signal upon amplification with a DNA band of 210 bases pairs, as well as 5 blood samples and 2 bone marrow specimens, with high sensitivity (10 picograms of Histoplasma capsulatum DNA) and specificity (no cross reactivity with P. brasiliensis, C. glabrata, C. neoformans and C. immitis). This technique has proved to be useful as a diagnostic technique. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Triton ENVELOPE(-55.615,-55.615,49.517,49.517) |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English Spanish |
topic |
histoplasmosis PCR ADN Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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histoplasmosis PCR ADN Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 María Teresa Colella Sofía Mata Claudia Hartung Celina Pérez Arantza Roselló Carolina Olaizola Carmen Fernández Silvia Magaldi Félix Toro Detection of Histoplasma Capsulatum by Nested-PCR in Clinical Specimens |
topic_facet |
histoplasmosis PCR ADN Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Histoplasmosis is a systemic mycosis whose diagnosis is based on clinical, mycological and immunological criteria. Identification of its etiological agent in biological samples is difficult to perform. We evaluated a nested-PCR technique to identify Histoplasma capsulatum in clinical specimens (blood and bone marrow) and reference strains. Three different methods for DNA isolation were tested based on the reagents Guanidine thiocyanate, Benzoyl chloride and Triton X-100. Among these procedures, Guanidine thiocyanate-based method yielded by utmost good qualityDNAand was easy to perform when used with clinical specimens. However, a better recovery of DNA was accomplished with the Triton-X-100 method. Reference strains of Paracoccidioides brasiliensis, Candida glabrata, Cryptococcus neoformans and Coccidioides immitis (C posadasii) were tested to assess PCR assay specificity. The samples were amplified by nested-PCR and analyzed by 3% agarose gel electrophoresis. Each of the reference strains showed the same signal upon amplification with a DNA band of 210 bases pairs, as well as 5 blood samples and 2 bone marrow specimens, with high sensitivity (10 picograms of Histoplasma capsulatum DNA) and specificity (no cross reactivity with P. brasiliensis, C. glabrata, C. neoformans and C. immitis). This technique has proved to be useful as a diagnostic technique. |
format |
Article in Journal/Newspaper |
author |
María Teresa Colella Sofía Mata Claudia Hartung Celina Pérez Arantza Roselló Carolina Olaizola Carmen Fernández Silvia Magaldi Félix Toro |
author_facet |
María Teresa Colella Sofía Mata Claudia Hartung Celina Pérez Arantza Roselló Carolina Olaizola Carmen Fernández Silvia Magaldi Félix Toro |
author_sort |
María Teresa Colella |
title |
Detection of Histoplasma Capsulatum by Nested-PCR in Clinical Specimens |
title_short |
Detection of Histoplasma Capsulatum by Nested-PCR in Clinical Specimens |
title_full |
Detection of Histoplasma Capsulatum by Nested-PCR in Clinical Specimens |
title_fullStr |
Detection of Histoplasma Capsulatum by Nested-PCR in Clinical Specimens |
title_full_unstemmed |
Detection of Histoplasma Capsulatum by Nested-PCR in Clinical Specimens |
title_sort |
detection of histoplasma capsulatum by nested-pcr in clinical specimens |
publisher |
Universidad del Zulia,Facultad de Medicina,Departamento de Enfermedades Infecciosas y Tropicales |
publishDate |
2007 |
url |
https://doaj.org/article/0d31ff9116a94e549776ef6d2b4a1900 |
long_lat |
ENVELOPE(-55.615,-55.615,49.517,49.517) |
geographic |
Arctic Triton |
geographic_facet |
Arctic Triton |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Kasmera, Vol 35, Iss 2, Pp 156-163 (2007) |
op_relation |
https://produccioncientificaluz.org/index.php/kasmera/article/view/4794 https://doaj.org/toc/0075-5222 https://doaj.org/toc/2477-9628 0075-5222 2477-9628 https://doaj.org/article/0d31ff9116a94e549776ef6d2b4a1900 |
_version_ |
1778521455179333632 |