Comparison of quantitative techniques including Xpert MTB/RIF to evaluate mycobacterial burden

 

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dc.contributor.author van Zyl-Smit, Richard N en_ZA
dc.contributor.author Binder, Anke en_ZA
dc.contributor.author Meldau, Richard en_ZA
dc.contributor.author Mishra, Hridesh en_ZA
dc.contributor.author Semple, Patricia L en_ZA
dc.contributor.author Theron, Grant en_ZA
dc.contributor.author Peter, Jonathan en_ZA
dc.contributor.author Whitelaw, Andrew en_ZA
dc.contributor.author Sharma, Suren K en_ZA
dc.contributor.author Warren, Robin en_ZA
dc.contributor.author Bateman, Eric D en_ZA
dc.contributor.author Dheda, Keertan en_ZA
dc.date.accessioned 2016-01-11T06:51:36Z
dc.date.available 2016-01-11T06:51:36Z
dc.date.issued 2011 en_ZA
dc.identifier.citation van Zyl-Smit, R. N., Binder, A., Meldau, R., Mishra, H., Semple, P. L., Theron, G., ... & Bateman, E. D. (2011). Comparison of quantitative techniques including Xpert MTB/RIF to evaluate mycobacterial burden. PLoS One, 6(12), e28815. doi:10.1371/journal.pone.0028815 en_ZA
dc.identifier.uri http://hdl.handle.net/11427/16248
dc.identifier.uri http://dx.doi.org/10.1371/journal.pone.0028815
dc.description.abstract Introduction: Accurate quantification of mycobacterial load is important for the evaluation of patient infectiousness, disease severity and monitoring treatment response in human and in-vitro laboratory models of disease. We hypothesized that newer techniques would perform as well as solid media culture to quantify mycobacterial burden in laboratory specimens. METHODS: We compared the turn-around-time, detection-threshold, dynamic range, reproducibility, relative discriminative ability, of 4 mycobacterial load determination techniques: automated liquid culture (BACTEC-MGIT-960), [ 3 H]-uracil incorporation assays, luciferase-reporter construct bioluminescence, and quantitative PCR(Xpert -MTB/RIF) using serial dilutions of Mycobacterium bovis and Mycobacterium tuberculosis H37RV. Mycobacterial colony-forming-units(CFU) using 7H10-Middlebrook solid media served as the reference standard. RESULTS: All 4 assays correlated well with the reference standard, however, bioluminescence and uracil assays had a detection threshold ≥1×10 3 organisms. By contrast, BACTEC-MGIT-960 liquid culture, although only providing results in days, was user-friendly, had the lowest detection threshold (<10 organisms), the greatest discriminative ability (1 vs. 10 organisms; p = 0.02), and the best reproducibility (coefficient of variance of 2% vs. 38% compared to uracil incorporation; p = 0.02). Xpert-MTB/RIF correlated well with mycobacterial load, had a rapid turn-around-time (<2 hours), was user friendly, but had a detection limit of ∼100 organisms. CONCLUSIONS: Choosing a technique to quantify mycobacterial burden for laboratory or clinical research depends on availability of resources and the question being addressed. Automated liquid culture has good discriminative ability and low detection threshold but results are only obtained in days. Xpert MTB/RIF provides rapid quantification of mycobacterial burden, but has a poorer discrimination and detection threshold. en_ZA
dc.language.iso eng en_ZA
dc.publisher Public Library of Science en_ZA
dc.rights This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. en_ZA
dc.rights.uri http://creativecommons.org/licenses/by/4.0 en_ZA
dc.source PLoS One en_ZA
dc.source.uri http://journals.plos.org/plosone en_ZA
dc.subject.other Mycobacterium tuberculosis en_ZA
dc.subject.other Uracils en_ZA
dc.subject.other Polymerase chain reaction en_ZA
dc.subject.other Liquids en_ZA
dc.subject.other Bioluminescence en_ZA
dc.subject.other Luminescence en_ZA
dc.subject.other Clinical laboratories en_ZA
dc.subject.other Tuberculosis en_ZA
dc.title Comparison of quantitative techniques including Xpert MTB/RIF to evaluate mycobacterial burden en_ZA
dc.type Journal Article en_ZA
dc.rights.holder © 2011 van Zyl-Smit et al en_ZA
uct.type.publication Research en_ZA
uct.type.resource Article en_ZA
dc.publisher.institution University of Cape Town
dc.publisher.faculty Faculty of Health Sciences en_ZA
dc.publisher.department Division of Pulmonology en_ZA
uct.type.filetype Text
uct.type.filetype Image


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This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Except where otherwise noted, this item's license is described as This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.