Performance of an in-house real-time polymerase chain reaction for identification of Mycobacterium tuberculosis isolates in laboratory routine diagnosis from a high burden setting

Abstract Brazil is one of the high burden countries for tuberculosis, and a rapid diagnosis is essential for effective control of the disease. In the present study, an in-house real-time polymerase chain reaction (PCR) assay targeting the mpt64 gene for identification of Mycobacterium tuberculosis complex isolates was evaluated under routine diagnosis conditions in a reference laboratory. From May 2011 to July 2012, 1,520 isolates of mycobacteria were prospectively submitted for phenotypic and/or PRA-hsp65 identification and to real-time PCR. The mpt64 real-time PCR showed 99.7% sensitivity and 96% specificity and detected 79.4% of the cases missed by phenotypic and PRA-hsp65 identification. The in-house real-time PCR assay showed high sensitivity and specificity and was successfully implemented in the routine diagnosis of tuberculosis in a reference laboratory from a high burden setting.

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Main Authors: Gallo,Juliana Failde, Pinhata,Juliana Maira Watanabe, Chimara,Erica, Gonçalves,Maria Gisele, Fukasawa,Lucila Okuyama, Oliveira,Rosangela Siqueira de
Format: Digital revista
Language:English
Published: Instituto Oswaldo Cruz, Ministério da Saúde 2016
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762016000900545
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spelling oai:scielo:S0074-027620160009005452016-09-15Performance of an in-house real-time polymerase chain reaction for identification of Mycobacterium tuberculosis isolates in laboratory routine diagnosis from a high burden settingGallo,Juliana FaildePinhata,Juliana Maira WatanabeChimara,EricaGonçalves,Maria GiseleFukasawa,Lucila OkuyamaOliveira,Rosangela Siqueira de molecular diagnostics MPT64 protein Mycobacterium tuberculosis real-time polymerase chain reaction tuberculosis Abstract Brazil is one of the high burden countries for tuberculosis, and a rapid diagnosis is essential for effective control of the disease. In the present study, an in-house real-time polymerase chain reaction (PCR) assay targeting the mpt64 gene for identification of Mycobacterium tuberculosis complex isolates was evaluated under routine diagnosis conditions in a reference laboratory. From May 2011 to July 2012, 1,520 isolates of mycobacteria were prospectively submitted for phenotypic and/or PRA-hsp65 identification and to real-time PCR. The mpt64 real-time PCR showed 99.7% sensitivity and 96% specificity and detected 79.4% of the cases missed by phenotypic and PRA-hsp65 identification. The in-house real-time PCR assay showed high sensitivity and specificity and was successfully implemented in the routine diagnosis of tuberculosis in a reference laboratory from a high burden setting.info:eu-repo/semantics/openAccessInstituto Oswaldo Cruz, Ministério da SaúdeMemórias do Instituto Oswaldo Cruz v.111 n.9 20162016-09-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762016000900545en10.1590/0074-02760160048
institution SCIELO
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country Brasil
countrycode BR
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region America del Sur
libraryname SciELO
language English
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author Gallo,Juliana Failde
Pinhata,Juliana Maira Watanabe
Chimara,Erica
Gonçalves,Maria Gisele
Fukasawa,Lucila Okuyama
Oliveira,Rosangela Siqueira de
spellingShingle Gallo,Juliana Failde
Pinhata,Juliana Maira Watanabe
Chimara,Erica
Gonçalves,Maria Gisele
Fukasawa,Lucila Okuyama
Oliveira,Rosangela Siqueira de
Performance of an in-house real-time polymerase chain reaction for identification of Mycobacterium tuberculosis isolates in laboratory routine diagnosis from a high burden setting
author_facet Gallo,Juliana Failde
Pinhata,Juliana Maira Watanabe
Chimara,Erica
Gonçalves,Maria Gisele
Fukasawa,Lucila Okuyama
Oliveira,Rosangela Siqueira de
author_sort Gallo,Juliana Failde
title Performance of an in-house real-time polymerase chain reaction for identification of Mycobacterium tuberculosis isolates in laboratory routine diagnosis from a high burden setting
title_short Performance of an in-house real-time polymerase chain reaction for identification of Mycobacterium tuberculosis isolates in laboratory routine diagnosis from a high burden setting
title_full Performance of an in-house real-time polymerase chain reaction for identification of Mycobacterium tuberculosis isolates in laboratory routine diagnosis from a high burden setting
title_fullStr Performance of an in-house real-time polymerase chain reaction for identification of Mycobacterium tuberculosis isolates in laboratory routine diagnosis from a high burden setting
title_full_unstemmed Performance of an in-house real-time polymerase chain reaction for identification of Mycobacterium tuberculosis isolates in laboratory routine diagnosis from a high burden setting
title_sort performance of an in-house real-time polymerase chain reaction for identification of mycobacterium tuberculosis isolates in laboratory routine diagnosis from a high burden setting
description Abstract Brazil is one of the high burden countries for tuberculosis, and a rapid diagnosis is essential for effective control of the disease. In the present study, an in-house real-time polymerase chain reaction (PCR) assay targeting the mpt64 gene for identification of Mycobacterium tuberculosis complex isolates was evaluated under routine diagnosis conditions in a reference laboratory. From May 2011 to July 2012, 1,520 isolates of mycobacteria were prospectively submitted for phenotypic and/or PRA-hsp65 identification and to real-time PCR. The mpt64 real-time PCR showed 99.7% sensitivity and 96% specificity and detected 79.4% of the cases missed by phenotypic and PRA-hsp65 identification. The in-house real-time PCR assay showed high sensitivity and specificity and was successfully implemented in the routine diagnosis of tuberculosis in a reference laboratory from a high burden setting.
publisher Instituto Oswaldo Cruz, Ministério da Saúde
publishDate 2016
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762016000900545
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