Application of PCR in serum samples for diagnosis of paracoccidioidomycosis in the southern Bahia-Brazil.
Paracoccidioidomycosis (PCM) cannot always be diagnosed by conventional means such as direct examination of histopathology or clinical samples, and serological methods, used as an alternative, still have many cases of cross-reactivity. In this scenario, molecular techniques seem to arise as a rapid...
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ftdoajarticles:oai:doaj.org/article:2e7a146adb414124b4fcd7181e1339e2 2023-05-15T15:10:52+02:00 Application of PCR in serum samples for diagnosis of paracoccidioidomycosis in the southern Bahia-Brazil. Lucas Dias Leila Falcão de Carvalho Carla C Romano 2012-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0001909 https://doaj.org/article/2e7a146adb414124b4fcd7181e1339e2 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3510084?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0001909 https://doaj.org/article/2e7a146adb414124b4fcd7181e1339e2 PLoS Neglected Tropical Diseases, Vol 6, Iss 11, p e1909 (2012) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2012 ftdoajarticles https://doi.org/10.1371/journal.pntd.0001909 2022-12-31T13:05:13Z Paracoccidioidomycosis (PCM) cannot always be diagnosed by conventional means such as direct examination of histopathology or clinical samples, and serological methods, used as an alternative, still have many cases of cross-reactivity. In this scenario, molecular techniques seem to arise as a rapid approach, specific and direct that could be used in the diagnosis of this mycosis. In this study we analyzed 76 serum samples from patients in southern Bahia suspected of having paracoccidioidomycosis using a conventional PCR with primers for the ITS1 ribosomal DNA of P. brasiliensis. Of these 76 patients, 5 were positive for PCM by double immunodiffusion and/or direct examination and histopathology. To test specificity of PCR, we used human DNA and three isolates of P. lutzii (1578, 01 and ED01). Additionally, we analyzed by serial dilutions of DNA the limit of detection of the assay. The test of PCR proved specific, as only a 144 bp fragment of the three isolates of P. lutzii and no human DNA was amplified. Detection limit was 1.1 pg/µL of DNA. Despite the high detection limit and specificity of PCR none of the 76 serum samples were found positive by PCR, but a biopsy specimen obtained from one of the patients with PCM was positive. These results, albeit limited, show that PCR is not effective in detecting DNA of P. brasiliensis or P. lutzii in serum, but could perhaps be used with other types of clinical samples, especially in those instances in which conventional methods fail. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 6 11 e1909 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Lucas Dias Leila Falcão de Carvalho Carla C Romano Application of PCR in serum samples for diagnosis of paracoccidioidomycosis in the southern Bahia-Brazil. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Paracoccidioidomycosis (PCM) cannot always be diagnosed by conventional means such as direct examination of histopathology or clinical samples, and serological methods, used as an alternative, still have many cases of cross-reactivity. In this scenario, molecular techniques seem to arise as a rapid approach, specific and direct that could be used in the diagnosis of this mycosis. In this study we analyzed 76 serum samples from patients in southern Bahia suspected of having paracoccidioidomycosis using a conventional PCR with primers for the ITS1 ribosomal DNA of P. brasiliensis. Of these 76 patients, 5 were positive for PCM by double immunodiffusion and/or direct examination and histopathology. To test specificity of PCR, we used human DNA and three isolates of P. lutzii (1578, 01 and ED01). Additionally, we analyzed by serial dilutions of DNA the limit of detection of the assay. The test of PCR proved specific, as only a 144 bp fragment of the three isolates of P. lutzii and no human DNA was amplified. Detection limit was 1.1 pg/µL of DNA. Despite the high detection limit and specificity of PCR none of the 76 serum samples were found positive by PCR, but a biopsy specimen obtained from one of the patients with PCM was positive. These results, albeit limited, show that PCR is not effective in detecting DNA of P. brasiliensis or P. lutzii in serum, but could perhaps be used with other types of clinical samples, especially in those instances in which conventional methods fail. |
format |
Article in Journal/Newspaper |
author |
Lucas Dias Leila Falcão de Carvalho Carla C Romano |
author_facet |
Lucas Dias Leila Falcão de Carvalho Carla C Romano |
author_sort |
Lucas Dias |
title |
Application of PCR in serum samples for diagnosis of paracoccidioidomycosis in the southern Bahia-Brazil. |
title_short |
Application of PCR in serum samples for diagnosis of paracoccidioidomycosis in the southern Bahia-Brazil. |
title_full |
Application of PCR in serum samples for diagnosis of paracoccidioidomycosis in the southern Bahia-Brazil. |
title_fullStr |
Application of PCR in serum samples for diagnosis of paracoccidioidomycosis in the southern Bahia-Brazil. |
title_full_unstemmed |
Application of PCR in serum samples for diagnosis of paracoccidioidomycosis in the southern Bahia-Brazil. |
title_sort |
application of pcr in serum samples for diagnosis of paracoccidioidomycosis in the southern bahia-brazil. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2012 |
url |
https://doi.org/10.1371/journal.pntd.0001909 https://doaj.org/article/2e7a146adb414124b4fcd7181e1339e2 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 6, Iss 11, p e1909 (2012) |
op_relation |
http://europepmc.org/articles/PMC3510084?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0001909 https://doaj.org/article/2e7a146adb414124b4fcd7181e1339e2 |
op_doi |
https://doi.org/10.1371/journal.pntd.0001909 |
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PLoS Neglected Tropical Diseases |
container_volume |
6 |
container_issue |
11 |
container_start_page |
e1909 |
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1766341808259858432 |