Evaluation of the modified colorimetric resazurin microtiter plate-based antibacterial assay for rapid and reliable tuberculosis drug susceptibility testing

Hdl Handle:
http://hdl.handle.net/10144/332873
Title:
Evaluation of the modified colorimetric resazurin microtiter plate-based antibacterial assay for rapid and reliable tuberculosis drug susceptibility testing
Authors:
Katawera, V; Siedner, M; Boum Ii, Y
Journal:
BMC Microbiology
Abstract:
BackgroundThe resazurin microtiter assay (classic REMA), a colorimetric liquid culture-based drug susceptibility assay for Mycobacterium tuberculosis (MTB), has been endorsed by the World Health Organization. The assay requires 8-16 days to obtain results, delaying management of drug resistant tuberculosis patients. A modified REMA which allows results in as little as 24 hours for bacterial strains, has been developed and validated using Staphylococcus aureus, but has not yet been evaluated for MTB. Therefore we assessed the performance of the modified REMA for rifampicin (RIF) and isoniazid (INH) susceptibility, using the classic REMA as the reference standard. We also compared simplicity (from the technicians¿ point of view), time taken to obtain results (rank-sum testing), specificity and Kappa statistics of the two methods.ResultsThe modified REMA, which is a one-step procedure, was found to be simpler to perform and results were obtained in a significantly shorter time (5 versus 9 days, p¿<¿0.0001) compared to the classic REMA due to addition of indicator and strain at the same time. The specificity of the modified REMA was low {46.8% (35.5% - 58.4%) for RIF and 13.9% (7.2% - 23.5%) for INH}. Kappa statistics were 16.0% for RIF and 2.0% for INH. Low specificity and kappa statistics are due to indicator reduction by the strains before complete drug activity.ConclusionAlthough modified REMA is faster and simpler compared to classic REMA, it is not reliable for MTB drug susceptibility testing.
Publisher:
BioMed Central
Issue Date:
7-Oct-2014
URI:
http://hdl.handle.net/10144/332873
DOI:
10.1186/s12866-014-0259-6
PubMed ID:
25287132
Language:
en
ISSN:
1471-2180
Appears in Collections:
TB

Full metadata record

DC FieldValue Language
dc.contributor.authorKatawera, Ven_GB
dc.contributor.authorSiedner, Men_GB
dc.contributor.authorBoum Ii, Yen_GB
dc.date.accessioned2014-10-17T15:40:16Z-
dc.date.available2014-10-17T15:40:16Z-
dc.date.issued2014-10-07-
dc.identifier.citationEvaluation of the modified colorimetric resazurin microtiter plate-based antibacterial assay for rapid and reliable tuberculosis drug susceptibility testing. 2014, 14 (1):259 BMC Microbiol.en_GB
dc.identifier.issn1471-2180-
dc.identifier.pmid25287132-
dc.identifier.doi10.1186/s12866-014-0259-6-
dc.identifier.urihttp://hdl.handle.net/10144/332873-
dc.description.abstractBackgroundThe resazurin microtiter assay (classic REMA), a colorimetric liquid culture-based drug susceptibility assay for Mycobacterium tuberculosis (MTB), has been endorsed by the World Health Organization. The assay requires 8-16 days to obtain results, delaying management of drug resistant tuberculosis patients. A modified REMA which allows results in as little as 24 hours for bacterial strains, has been developed and validated using Staphylococcus aureus, but has not yet been evaluated for MTB. Therefore we assessed the performance of the modified REMA for rifampicin (RIF) and isoniazid (INH) susceptibility, using the classic REMA as the reference standard. We also compared simplicity (from the technicians¿ point of view), time taken to obtain results (rank-sum testing), specificity and Kappa statistics of the two methods.ResultsThe modified REMA, which is a one-step procedure, was found to be simpler to perform and results were obtained in a significantly shorter time (5 versus 9 days, p¿<¿0.0001) compared to the classic REMA due to addition of indicator and strain at the same time. The specificity of the modified REMA was low {46.8% (35.5% - 58.4%) for RIF and 13.9% (7.2% - 23.5%) for INH}. Kappa statistics were 16.0% for RIF and 2.0% for INH. Low specificity and kappa statistics are due to indicator reduction by the strains before complete drug activity.ConclusionAlthough modified REMA is faster and simpler compared to classic REMA, it is not reliable for MTB drug susceptibility testing.en_GB
dc.languageENG-
dc.language.isoenen
dc.publisherBioMed Centralen_GB
dc.rightsArchived with thanks to BMC Microbiologyen_GB
dc.titleEvaluation of the modified colorimetric resazurin microtiter plate-based antibacterial assay for rapid and reliable tuberculosis drug susceptibility testingen
dc.identifier.journalBMC Microbiologyen_GB
All Items in MSF are protected by copyright, with all rights reserved, unless otherwise indicated.