Validation of a real-time PCR assay for the molecular identification of Mycobacterium tuberculosis

Detalhes bibliográficos
Autor(a) principal: Sales,Mariana L.
Data de Publicação: 2014
Outros Autores: 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
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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spelling 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
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