Comparison of two laboratory-developed PCR methods for the diagnosis of pulmonary tuberculosis in Brazilian patients with and without HIV infection
Autor(a) principal: | |
---|---|
Data de Publicação: | 2011 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFRGS |
Texto Completo: | http://hdl.handle.net/10183/30950 |
Resumo: | Background: Direct smear examination with Ziehl-Neelsen (ZN) staining for the diagnosis of pulmonary tuberculosis (PTB) is cheap and easy to use, but its low sensitivity is a major drawback, particularly in HIV seropositive patients. As such, new tools for laboratory diagnosis are urgently needed to improve the case detection rate, especially in regions with a high prevalence of TB and HIV. Objective: To evaluate the performance of two in house PCR (Polymerase Chain Reaction): PCR dot-blot methodology (PCR dot-blot) and PCR agarose gel electrophoresis (PCR-AG) for the diagnosis of Pulmonary Tuberculosis (PTB) in HIV seropositive and HIV seronegative patients. Methods: A prospective study was conducted (from May 2003 to May 2004) in a TB/HIV reference hospital. Sputum specimens from 277 PTB suspects were tested by Acid Fast Bacilli (AFB) smear, Culture and in house PCR assays (PCR dot-blot and PCR-AG) and their performances evaluated. Positive cultures combined with the definition of clinical pulmonary TB were employed as the gold standard. Results: The overall prevalence of PTB was 46% (128/277); in HIV+, prevalence was 54.0% (40/74). The sensitivity and specificity of PCR dot-blot were 74% (CI 95%; 66.1%-81.2%) and 85% (CI 95%; 78.8%-90.3%); and of PCR-AG were 43% (CI 95%; 34.5%-51.6%) and 76% (CI 95%; 69.2%-82.8%), respectively. For HIV seropositive and HIV seronegative samples, sensitivities of PCR dot-blot (72% vs 75%; p = 0.46) and PCR-AG (42% vs 43%; p = 0.54) were similar. Among HIV seronegative patients and PTB suspects, ROC analysis presented the following values for the AFB smear (0.837), Culture (0.926), PCR dot-blot (0.801) and PCR-AG (0.599). In HIV seropositive patients, these area values were (0.713), (0.900), (0.789) and (0.595), respectively. Conclusion: Results of this study demonstrate that the in house PCR dot blot may be an improvement for ruling out PTB diagnosis in PTB suspects assisted at hospitals with a high prevalence of TB/HIV. |
id |
UFRGS-2_a53c3656124b01ab3554dc02711164d6 |
---|---|
oai_identifier_str |
oai:www.lume.ufrgs.br:10183/30950 |
network_acronym_str |
UFRGS-2 |
network_name_str |
Repositório Institucional da UFRGS |
repository_id_str |
|
spelling |
Scherer, Luciene CardosoSperhacke, Rosa DeaJarczewski, Carla AdrianeCafrune, Patricia IzquierdoMichelon, Candice TosiRuppenthal, Rubia DeniseRibeiro, Marta OsórioRuffino-Netto, AntonioRossetti, Maria Lucia RosaKritski, Afrânio Lineu2011-08-09T06:01:06Z20111471-2466http://hdl.handle.net/10183/30950000782096Background: Direct smear examination with Ziehl-Neelsen (ZN) staining for the diagnosis of pulmonary tuberculosis (PTB) is cheap and easy to use, but its low sensitivity is a major drawback, particularly in HIV seropositive patients. As such, new tools for laboratory diagnosis are urgently needed to improve the case detection rate, especially in regions with a high prevalence of TB and HIV. Objective: To evaluate the performance of two in house PCR (Polymerase Chain Reaction): PCR dot-blot methodology (PCR dot-blot) and PCR agarose gel electrophoresis (PCR-AG) for the diagnosis of Pulmonary Tuberculosis (PTB) in HIV seropositive and HIV seronegative patients. Methods: A prospective study was conducted (from May 2003 to May 2004) in a TB/HIV reference hospital. Sputum specimens from 277 PTB suspects were tested by Acid Fast Bacilli (AFB) smear, Culture and in house PCR assays (PCR dot-blot and PCR-AG) and their performances evaluated. Positive cultures combined with the definition of clinical pulmonary TB were employed as the gold standard. Results: The overall prevalence of PTB was 46% (128/277); in HIV+, prevalence was 54.0% (40/74). The sensitivity and specificity of PCR dot-blot were 74% (CI 95%; 66.1%-81.2%) and 85% (CI 95%; 78.8%-90.3%); and of PCR-AG were 43% (CI 95%; 34.5%-51.6%) and 76% (CI 95%; 69.2%-82.8%), respectively. For HIV seropositive and HIV seronegative samples, sensitivities of PCR dot-blot (72% vs 75%; p = 0.46) and PCR-AG (42% vs 43%; p = 0.54) were similar. Among HIV seronegative patients and PTB suspects, ROC analysis presented the following values for the AFB smear (0.837), Culture (0.926), PCR dot-blot (0.801) and PCR-AG (0.599). In HIV seropositive patients, these area values were (0.713), (0.900), (0.789) and (0.595), respectively. Conclusion: Results of this study demonstrate that the in house PCR dot blot may be an improvement for ruling out PTB diagnosis in PTB suspects assisted at hospitals with a high prevalence of TB/HIV.application/pdfengBMC Pulmonary medicine. London. Vol, 11, no. 15 (29 mar. 2011), 10 p.Tuberculose pulmonarReação em cadeia da polimeraseHIVComparison of two laboratory-developed PCR methods for the diagnosis of pulmonary tuberculosis in Brazilian patients with and without HIV infectionEstrangeiroinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFRGSinstname:Universidade Federal do Rio Grande do Sul (UFRGS)instacron:UFRGSTEXT000782096.pdf.txt000782096.pdf.txtExtracted Texttext/plain43828http://www.lume.ufrgs.br/bitstream/10183/30950/2/000782096.pdf.txt56a91b85ff7cfec1e55f070855feb24bMD52ORIGINAL000782096.pdf000782096.pdfTexto completo (inglês)application/pdf243850http://www.lume.ufrgs.br/bitstream/10183/30950/1/000782096.pdf026341bff13e42a9999172b44c115f04MD51THUMBNAIL000782096.pdf.jpg000782096.pdf.jpgGenerated Thumbnailimage/jpeg1969http://www.lume.ufrgs.br/bitstream/10183/30950/3/000782096.pdf.jpg55e04cbffb02b303ea2bf757cbef94eeMD5310183/309502019-01-11 04:08:23.372488oai:www.lume.ufrgs.br:10183/30950Repositório de PublicaçõesPUBhttps://lume.ufrgs.br/oai/requestopendoar:2019-01-11T06:08:23Repositório Institucional da UFRGS - Universidade Federal do Rio Grande do Sul (UFRGS)false |
dc.title.pt_BR.fl_str_mv |
Comparison of two laboratory-developed PCR methods for the diagnosis of pulmonary tuberculosis in Brazilian patients with and without HIV infection |
title |
Comparison of two laboratory-developed PCR methods for the diagnosis of pulmonary tuberculosis in Brazilian patients with and without HIV infection |
spellingShingle |
Comparison of two laboratory-developed PCR methods for the diagnosis of pulmonary tuberculosis in Brazilian patients with and without HIV infection Scherer, Luciene Cardoso Tuberculose pulmonar Reação em cadeia da polimerase HIV |
title_short |
Comparison of two laboratory-developed PCR methods for the diagnosis of pulmonary tuberculosis in Brazilian patients with and without HIV infection |
title_full |
Comparison of two laboratory-developed PCR methods for the diagnosis of pulmonary tuberculosis in Brazilian patients with and without HIV infection |
title_fullStr |
Comparison of two laboratory-developed PCR methods for the diagnosis of pulmonary tuberculosis in Brazilian patients with and without HIV infection |
title_full_unstemmed |
Comparison of two laboratory-developed PCR methods for the diagnosis of pulmonary tuberculosis in Brazilian patients with and without HIV infection |
title_sort |
Comparison of two laboratory-developed PCR methods for the diagnosis of pulmonary tuberculosis in Brazilian patients with and without HIV infection |
author |
Scherer, Luciene Cardoso |
author_facet |
Scherer, Luciene Cardoso Sperhacke, Rosa Dea Jarczewski, Carla Adriane Cafrune, Patricia Izquierdo Michelon, Candice Tosi Ruppenthal, Rubia Denise Ribeiro, Marta Osório Ruffino-Netto, Antonio Rossetti, Maria Lucia Rosa Kritski, Afrânio Lineu |
author_role |
author |
author2 |
Sperhacke, Rosa Dea Jarczewski, Carla Adriane Cafrune, Patricia Izquierdo Michelon, Candice Tosi Ruppenthal, Rubia Denise Ribeiro, Marta Osório Ruffino-Netto, Antonio Rossetti, Maria Lucia Rosa Kritski, Afrânio Lineu |
author2_role |
author author author author author author author author author |
dc.contributor.author.fl_str_mv |
Scherer, Luciene Cardoso Sperhacke, Rosa Dea Jarczewski, Carla Adriane Cafrune, Patricia Izquierdo Michelon, Candice Tosi Ruppenthal, Rubia Denise Ribeiro, Marta Osório Ruffino-Netto, Antonio Rossetti, Maria Lucia Rosa Kritski, Afrânio Lineu |
dc.subject.por.fl_str_mv |
Tuberculose pulmonar Reação em cadeia da polimerase HIV |
topic |
Tuberculose pulmonar Reação em cadeia da polimerase HIV |
description |
Background: Direct smear examination with Ziehl-Neelsen (ZN) staining for the diagnosis of pulmonary tuberculosis (PTB) is cheap and easy to use, but its low sensitivity is a major drawback, particularly in HIV seropositive patients. As such, new tools for laboratory diagnosis are urgently needed to improve the case detection rate, especially in regions with a high prevalence of TB and HIV. Objective: To evaluate the performance of two in house PCR (Polymerase Chain Reaction): PCR dot-blot methodology (PCR dot-blot) and PCR agarose gel electrophoresis (PCR-AG) for the diagnosis of Pulmonary Tuberculosis (PTB) in HIV seropositive and HIV seronegative patients. Methods: A prospective study was conducted (from May 2003 to May 2004) in a TB/HIV reference hospital. Sputum specimens from 277 PTB suspects were tested by Acid Fast Bacilli (AFB) smear, Culture and in house PCR assays (PCR dot-blot and PCR-AG) and their performances evaluated. Positive cultures combined with the definition of clinical pulmonary TB were employed as the gold standard. Results: The overall prevalence of PTB was 46% (128/277); in HIV+, prevalence was 54.0% (40/74). The sensitivity and specificity of PCR dot-blot were 74% (CI 95%; 66.1%-81.2%) and 85% (CI 95%; 78.8%-90.3%); and of PCR-AG were 43% (CI 95%; 34.5%-51.6%) and 76% (CI 95%; 69.2%-82.8%), respectively. For HIV seropositive and HIV seronegative samples, sensitivities of PCR dot-blot (72% vs 75%; p = 0.46) and PCR-AG (42% vs 43%; p = 0.54) were similar. Among HIV seronegative patients and PTB suspects, ROC analysis presented the following values for the AFB smear (0.837), Culture (0.926), PCR dot-blot (0.801) and PCR-AG (0.599). In HIV seropositive patients, these area values were (0.713), (0.900), (0.789) and (0.595), respectively. Conclusion: Results of this study demonstrate that the in house PCR dot blot may be an improvement for ruling out PTB diagnosis in PTB suspects assisted at hospitals with a high prevalence of TB/HIV. |
publishDate |
2011 |
dc.date.accessioned.fl_str_mv |
2011-08-09T06:01:06Z |
dc.date.issued.fl_str_mv |
2011 |
dc.type.driver.fl_str_mv |
Estrangeiro info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10183/30950 |
dc.identifier.issn.pt_BR.fl_str_mv |
1471-2466 |
dc.identifier.nrb.pt_BR.fl_str_mv |
000782096 |
identifier_str_mv |
1471-2466 000782096 |
url |
http://hdl.handle.net/10183/30950 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.pt_BR.fl_str_mv |
BMC Pulmonary medicine. London. Vol, 11, no. 15 (29 mar. 2011), 10 p. |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFRGS instname:Universidade Federal do Rio Grande do Sul (UFRGS) instacron:UFRGS |
instname_str |
Universidade Federal do Rio Grande do Sul (UFRGS) |
instacron_str |
UFRGS |
institution |
UFRGS |
reponame_str |
Repositório Institucional da UFRGS |
collection |
Repositório Institucional da UFRGS |
bitstream.url.fl_str_mv |
http://www.lume.ufrgs.br/bitstream/10183/30950/2/000782096.pdf.txt http://www.lume.ufrgs.br/bitstream/10183/30950/1/000782096.pdf http://www.lume.ufrgs.br/bitstream/10183/30950/3/000782096.pdf.jpg |
bitstream.checksum.fl_str_mv |
56a91b85ff7cfec1e55f070855feb24b 026341bff13e42a9999172b44c115f04 55e04cbffb02b303ea2bf757cbef94ee |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositório Institucional da UFRGS - Universidade Federal do Rio Grande do Sul (UFRGS) |
repository.mail.fl_str_mv |
|
_version_ |
1801224729676218368 |