Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Outros Autores: | , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Brazilian Journal of Microbiology |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000400029 |
Resumo: | Mycobacterium tuberculosis is the major cause of tuberculosis in humans. This bacillus gained prominence with the occurrence of HIV, presenting itself as an important opportunistic infection associated with acquired immunodeficiency syndrome (AIDS). The current study aimed to develop a real-time PCR using Eva Green technology for molecular identification of M. tuberculosis isolates. The primers were designed to Rv1510 gene. Ninety nine samples of M. tuberculosis and sixty samples of M. bovis were tested and no sample of the bovine bacillus was detected by the qPCR. Statistical tests showed no difference between the qPCR and biochemical tests used to identify the Mycobacterium tuberculosis. The correlation between tests was perfect with Kappa index of 1.0 (p < 0.001, CI = 0.84 - 1.0). The diagnostic sensitivity and specificity were 100% (CI = 95.94% - 100%) and 100% (CI = 93.98% - 100%). This qPCR was developed with the goal of diagnosing the bacillus M. tuberculosis in samples of bacterial suspension. TB reference laboratories (health and agriculture sectors), public health programs and epidemiological studies probably may benefit from such method. |
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Brazilian Journal of Microbiology |
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Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosisMycobacterium tuberculosisdiagnosisreal time PCRMycobacterium tuberculosis is the major cause of tuberculosis in humans. This bacillus gained prominence with the occurrence of HIV, presenting itself as an important opportunistic infection associated with acquired immunodeficiency syndrome (AIDS). The current study aimed to develop a real-time PCR using Eva Green technology for molecular identification of M. tuberculosis isolates. The primers were designed to Rv1510 gene. Ninety nine samples of M. tuberculosis and sixty samples of M. bovis were tested and no sample of the bovine bacillus was detected by the qPCR. Statistical tests showed no difference between the qPCR and biochemical tests used to identify the Mycobacterium tuberculosis. The correlation between tests was perfect with Kappa index of 1.0 (p < 0.001, CI = 0.84 - 1.0). The diagnostic sensitivity and specificity were 100% (CI = 95.94% - 100%) and 100% (CI = 93.98% - 100%). This qPCR was developed with the goal of diagnosing the bacillus M. tuberculosis in samples of bacterial suspension. TB reference laboratories (health and agriculture sectors), public health programs and epidemiological studies probably may benefit from such method.Sociedade Brasileira de Microbiologia2014-12-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000400029Brazilian Journal of Microbiology v.45 n.4 2014reponame:Brazilian Journal of Microbiologyinstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1590/S1517-83822014000400029info:eu-repo/semantics/openAccessSales,Mariana L.Fonseca Júnior,Antônio AugustoOrzil,LíviaAlencar,Andrea PadilhaSilva,Marcio RobertoIssa,Marina AzevedoSoares Filho,Paulo MartinsLage,Andrey PereiraHeinemann,Marcos Bryaneng2015-02-13T00:00:00Zoai:scielo:S1517-83822014000400029Revistahttps://www.scielo.br/j/bjm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||mbmartin@usp.br1678-44051517-8382opendoar:2015-02-13T00:00Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)false |
dc.title.none.fl_str_mv |
Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis |
title |
Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis |
spellingShingle |
Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis Sales,Mariana L. Mycobacterium tuberculosis diagnosis real time PCR |
title_short |
Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis |
title_full |
Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis |
title_fullStr |
Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis |
title_full_unstemmed |
Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis |
title_sort |
Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis |
author |
Sales,Mariana L. |
author_facet |
Sales,Mariana L. Fonseca Júnior,Antônio Augusto Orzil,Lívia Alencar,Andrea Padilha Silva,Marcio Roberto Issa,Marina Azevedo Soares Filho,Paulo Martins Lage,Andrey Pereira Heinemann,Marcos Bryan |
author_role |
author |
author2 |
Fonseca Júnior,Antônio Augusto Orzil,Lívia Alencar,Andrea Padilha Silva,Marcio Roberto Issa,Marina Azevedo Soares Filho,Paulo Martins Lage,Andrey Pereira Heinemann,Marcos Bryan |
author2_role |
author author author author author author author author |
dc.contributor.author.fl_str_mv |
Sales,Mariana L. Fonseca Júnior,Antônio Augusto Orzil,Lívia Alencar,Andrea Padilha Silva,Marcio Roberto Issa,Marina Azevedo Soares Filho,Paulo Martins Lage,Andrey Pereira Heinemann,Marcos Bryan |
dc.subject.por.fl_str_mv |
Mycobacterium tuberculosis diagnosis real time PCR |
topic |
Mycobacterium tuberculosis diagnosis real time PCR |
description |
Mycobacterium tuberculosis is the major cause of tuberculosis in humans. This bacillus gained prominence with the occurrence of HIV, presenting itself as an important opportunistic infection associated with acquired immunodeficiency syndrome (AIDS). The current study aimed to develop a real-time PCR using Eva Green technology for molecular identification of M. tuberculosis isolates. The primers were designed to Rv1510 gene. Ninety nine samples of M. tuberculosis and sixty samples of M. bovis were tested and no sample of the bovine bacillus was detected by the qPCR. Statistical tests showed no difference between the qPCR and biochemical tests used to identify the Mycobacterium tuberculosis. The correlation between tests was perfect with Kappa index of 1.0 (p < 0.001, CI = 0.84 - 1.0). The diagnostic sensitivity and specificity were 100% (CI = 95.94% - 100%) and 100% (CI = 93.98% - 100%). This qPCR was developed with the goal of diagnosing the bacillus M. tuberculosis in samples of bacterial suspension. TB reference laboratories (health and agriculture sectors), public health programs and epidemiological studies probably may benefit from such method. |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014-12-01 |
dc.type.driver.fl_str_mv |
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://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000400029 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000400029 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S1517-83822014000400029 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
dc.source.none.fl_str_mv |
Brazilian Journal of Microbiology v.45 n.4 2014 reponame:Brazilian Journal of Microbiology instname:Sociedade Brasileira de Microbiologia (SBM) instacron:SBM |
instname_str |
Sociedade Brasileira de Microbiologia (SBM) |
instacron_str |
SBM |
institution |
SBM |
reponame_str |
Brazilian Journal of Microbiology |
collection |
Brazilian Journal of Microbiology |
repository.name.fl_str_mv |
Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM) |
repository.mail.fl_str_mv |
bjm@sbmicrobiologia.org.br||mbmartin@usp.br |
_version_ |
1752122206942396416 |