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

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dc.contributor.authorvan Zyl-Smit, Richard Nen_ZA
dc.contributor.authorBinder, Ankeen_ZA
dc.contributor.authorMeldau, Richarden_ZA
dc.contributor.authorMishra, Hrideshen_ZA
dc.contributor.authorSemple, Patricia Len_ZA
dc.contributor.authorTheron, Granten_ZA
dc.contributor.authorPeter, Jonathanen_ZA
dc.contributor.authorWhitelaw, Andrewen_ZA
dc.contributor.authorSharma, Suren Ken_ZA
dc.contributor.authorWarren, Robinen_ZA
dc.contributor.authorBateman, Eric Den_ZA
dc.contributor.authorDheda, Keertanen_ZA
dc.date.accessioned2016-01-11T06:51:36Z
dc.date.available2016-01-11T06:51:36Z
dc.date.issued2011en_ZA
dc.description.abstractIntroduction: 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.identifier.apacitationvan Zyl-Smit, R. N., Binder, A., Meldau, R., Mishra, H., Semple, P. L., Theron, G., ... Dheda, K. (2011). Comparison of quantitative techniques including Xpert MTB/RIF to evaluate mycobacterial burden. <i>PLoS One</i>, http://hdl.handle.net/11427/16248en_ZA
dc.identifier.chicagocitationvan Zyl-Smit, Richard N, Anke Binder, Richard Meldau, Hridesh Mishra, Patricia L Semple, Grant Theron, Jonathan Peter, et al "Comparison of quantitative techniques including Xpert MTB/RIF to evaluate mycobacterial burden." <i>PLoS One</i> (2011) http://hdl.handle.net/11427/16248en_ZA
dc.identifier.citationvan 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.0028815en_ZA
dc.identifier.ris TY - Journal Article AU - van Zyl-Smit, Richard N AU - Binder, Anke AU - Meldau, Richard AU - Mishra, Hridesh AU - Semple, Patricia L AU - Theron, Grant AU - Peter, Jonathan AU - Whitelaw, Andrew AU - Sharma, Suren K AU - Warren, Robin AU - Bateman, Eric D AU - Dheda, Keertan AB - 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. DA - 2011 DB - OpenUCT DO - 10.1371/journal.pone.0028815 DP - University of Cape Town J1 - PLoS One LK - https://open.uct.ac.za PB - University of Cape Town PY - 2011 T1 - Comparison of quantitative techniques including Xpert MTB/RIF to evaluate mycobacterial burden TI - Comparison of quantitative techniques including Xpert MTB/RIF to evaluate mycobacterial burden UR - http://hdl.handle.net/11427/16248 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/16248
dc.identifier.urihttp://dx.doi.org/10.1371/journal.pone.0028815
dc.identifier.vancouvercitationvan Zyl-Smit RN, Binder A, Meldau R, Mishra H, Semple PL, Theron G, et al. Comparison of quantitative techniques including Xpert MTB/RIF to evaluate mycobacterial burden. PLoS One. 2011; http://hdl.handle.net/11427/16248.en_ZA
dc.language.isoengen_ZA
dc.publisherPublic Library of Scienceen_ZA
dc.publisher.departmentDivision of Pulmonologyen_ZA
dc.publisher.facultyFaculty of Health Sciencesen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.rightsThis 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.holder© 2011 van Zyl-Smit et alen_ZA
dc.rights.urihttp://creativecommons.org/licenses/by/4.0en_ZA
dc.sourcePLoS Oneen_ZA
dc.source.urihttp://journals.plos.org/plosoneen_ZA
dc.subject.otherMycobacterium tuberculosisen_ZA
dc.subject.otherUracilsen_ZA
dc.subject.otherPolymerase chain reactionen_ZA
dc.subject.otherLiquidsen_ZA
dc.subject.otherBioluminescenceen_ZA
dc.subject.otherLuminescenceen_ZA
dc.subject.otherClinical laboratoriesen_ZA
dc.subject.otherTuberculosisen_ZA
dc.titleComparison of quantitative techniques including Xpert MTB/RIF to evaluate mycobacterial burdenen_ZA
dc.typeJournal Articleen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
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