Combination of commercially available molecular assays and culture based methods in diagnosis of tuberculosis and drug resistant tuberculosis

ABSTRACT Early diagnosis of tuberculosis is of major clinical importance. Among 4733 clinical specimens collected from 3363 patients and subjected to Ziehl-Neelsen microscopy, 4109 were inoculated onto Löwenstein-Jensen slants and 3139 in Bactec/9000MB. Polymerase Chain Reaction (PCR) was performed in 3139 specimens, whereas, a genotypic assay was directly applied in 93 Mycobacterium tuberculosis complex PCR-positive for isoniazid and rifampicin resistance detection specimens (GenoType MTBDRplus). Recovered M. tuberculosis isolates (64) as well as, 21 more sent from Regional Hospitals were tested for antimycobacterial resistance with a phenotypic (manual MGIT-SIRE) and a genotypic assay (GenoType MTBDRplus). PCR in the clinical specimens showed excellent specificity (97.4%) and accuracy (96.8%), good sensitivity (70.4%), but low positive predictive value (40.3%). MGIT-SIRE performed to M. tuberculosis did not confer a reliable result in 16 isolates. Of the remaining 69 isolates, 15 were resistant to streptomycin, seven to isoniazid, seven to ethambutol and five to rifampicin. GenoType MTBDRplus correctly detected isoniazid (seven) and rifampicin-resistant M. tuberculosis strains (five), showing an excellent performance overall (100%). Susceptibility results by the molecular assay applied directly to clinical specimens were identical to those obtained from recovered isolates of the corresponding patients. Combining molecular and conventional methods greatly contribute to early diagnosis and accurate susceptibility testing of tuberculosis.

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Main Authors: Gkaravela,Lamprini, Papadimitriou-Olivgeris,Matthaios, Foka,Antigoni, Kolonitsiou,Fevronia, Spiliopoulou,Anastasia, Charokopos,Nikolaos, Voulgaridis,Apostolos, Tsiamita,Maria, Marangos,Markos, Anastassiou,Evangelos D., Spiliopoulou,Iris
Format: Digital revista
Language:English
Published: Sociedade Brasileira de Microbiologia 2017
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000400785
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spelling oai:scielo:S1517-838220170004007852017-10-31Combination of commercially available molecular assays and culture based methods in diagnosis of tuberculosis and drug resistant tuberculosisGkaravela,LampriniPapadimitriou-Olivgeris,MatthaiosFoka,AntigoniKolonitsiou,FevroniaSpiliopoulou,AnastasiaCharokopos,NikolaosVoulgaridis,ApostolosTsiamita,MariaMarangos,MarkosAnastassiou,Evangelos D.Spiliopoulou,Iris Mycobacterium tuberculosis PCR GenoType MTBDRplus ABSTRACT Early diagnosis of tuberculosis is of major clinical importance. Among 4733 clinical specimens collected from 3363 patients and subjected to Ziehl-Neelsen microscopy, 4109 were inoculated onto Löwenstein-Jensen slants and 3139 in Bactec/9000MB. Polymerase Chain Reaction (PCR) was performed in 3139 specimens, whereas, a genotypic assay was directly applied in 93 Mycobacterium tuberculosis complex PCR-positive for isoniazid and rifampicin resistance detection specimens (GenoType MTBDRplus). Recovered M. tuberculosis isolates (64) as well as, 21 more sent from Regional Hospitals were tested for antimycobacterial resistance with a phenotypic (manual MGIT-SIRE) and a genotypic assay (GenoType MTBDRplus). PCR in the clinical specimens showed excellent specificity (97.4%) and accuracy (96.8%), good sensitivity (70.4%), but low positive predictive value (40.3%). MGIT-SIRE performed to M. tuberculosis did not confer a reliable result in 16 isolates. Of the remaining 69 isolates, 15 were resistant to streptomycin, seven to isoniazid, seven to ethambutol and five to rifampicin. GenoType MTBDRplus correctly detected isoniazid (seven) and rifampicin-resistant M. tuberculosis strains (five), showing an excellent performance overall (100%). Susceptibility results by the molecular assay applied directly to clinical specimens were identical to those obtained from recovered isolates of the corresponding patients. Combining molecular and conventional methods greatly contribute to early diagnosis and accurate susceptibility testing of tuberculosis.info:eu-repo/semantics/openAccessSociedade Brasileira de MicrobiologiaBrazilian Journal of Microbiology v.48 n.4 20172017-12-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000400785en10.1016/j.bjm.2017.04.001
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libraryname SciELO
language English
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author Gkaravela,Lamprini
Papadimitriou-Olivgeris,Matthaios
Foka,Antigoni
Kolonitsiou,Fevronia
Spiliopoulou,Anastasia
Charokopos,Nikolaos
Voulgaridis,Apostolos
Tsiamita,Maria
Marangos,Markos
Anastassiou,Evangelos D.
Spiliopoulou,Iris
spellingShingle Gkaravela,Lamprini
Papadimitriou-Olivgeris,Matthaios
Foka,Antigoni
Kolonitsiou,Fevronia
Spiliopoulou,Anastasia
Charokopos,Nikolaos
Voulgaridis,Apostolos
Tsiamita,Maria
Marangos,Markos
Anastassiou,Evangelos D.
Spiliopoulou,Iris
Combination of commercially available molecular assays and culture based methods in diagnosis of tuberculosis and drug resistant tuberculosis
author_facet Gkaravela,Lamprini
Papadimitriou-Olivgeris,Matthaios
Foka,Antigoni
Kolonitsiou,Fevronia
Spiliopoulou,Anastasia
Charokopos,Nikolaos
Voulgaridis,Apostolos
Tsiamita,Maria
Marangos,Markos
Anastassiou,Evangelos D.
Spiliopoulou,Iris
author_sort Gkaravela,Lamprini
title Combination of commercially available molecular assays and culture based methods in diagnosis of tuberculosis and drug resistant tuberculosis
title_short Combination of commercially available molecular assays and culture based methods in diagnosis of tuberculosis and drug resistant tuberculosis
title_full Combination of commercially available molecular assays and culture based methods in diagnosis of tuberculosis and drug resistant tuberculosis
title_fullStr Combination of commercially available molecular assays and culture based methods in diagnosis of tuberculosis and drug resistant tuberculosis
title_full_unstemmed Combination of commercially available molecular assays and culture based methods in diagnosis of tuberculosis and drug resistant tuberculosis
title_sort combination of commercially available molecular assays and culture based methods in diagnosis of tuberculosis and drug resistant tuberculosis
description ABSTRACT Early diagnosis of tuberculosis is of major clinical importance. Among 4733 clinical specimens collected from 3363 patients and subjected to Ziehl-Neelsen microscopy, 4109 were inoculated onto Löwenstein-Jensen slants and 3139 in Bactec/9000MB. Polymerase Chain Reaction (PCR) was performed in 3139 specimens, whereas, a genotypic assay was directly applied in 93 Mycobacterium tuberculosis complex PCR-positive for isoniazid and rifampicin resistance detection specimens (GenoType MTBDRplus). Recovered M. tuberculosis isolates (64) as well as, 21 more sent from Regional Hospitals were tested for antimycobacterial resistance with a phenotypic (manual MGIT-SIRE) and a genotypic assay (GenoType MTBDRplus). PCR in the clinical specimens showed excellent specificity (97.4%) and accuracy (96.8%), good sensitivity (70.4%), but low positive predictive value (40.3%). MGIT-SIRE performed to M. tuberculosis did not confer a reliable result in 16 isolates. Of the remaining 69 isolates, 15 were resistant to streptomycin, seven to isoniazid, seven to ethambutol and five to rifampicin. GenoType MTBDRplus correctly detected isoniazid (seven) and rifampicin-resistant M. tuberculosis strains (five), showing an excellent performance overall (100%). Susceptibility results by the molecular assay applied directly to clinical specimens were identical to those obtained from recovered isolates of the corresponding patients. Combining molecular and conventional methods greatly contribute to early diagnosis and accurate susceptibility testing of tuberculosis.
publisher Sociedade Brasileira de Microbiologia
publishDate 2017
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000400785
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