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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
Comparison of quantitative techniques including Xpert MTB/RIF to evaluate mycobacterial burden
PLoS ONE, Volume 6, No. 12, Article e28815, Year 2011
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Description
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), [ 3H]-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. © 2011 van Zyl-Smit et al.
Authors & Co-Authors
van Zyl-Smit, Richard N.
South Africa, Cape Town
University of Cape Town Lung Institute
Binder, Anke B.
South Africa, Cape Town
University of Cape Town Lung Institute
Meldau, Richard
South Africa, Cape Town
University of Cape Town Lung Institute
Mishra, Hridesh Kumar
India, New Delhi
All India Institute of Medical Sciences, new Delhi
Semple, Patricia L.
South Africa, Cape Town
University of Cape Town Lung Institute
Theron, Grant
South Africa, Cape Town
University of Cape Town Lung Institute
Peter, Jonathan Grant
South Africa, Cape Town
University of Cape Town Lung Institute
Whitelaw, Andrew C.
South Africa, Cape Town
University of Cape Town
Sharma, Surendra Kumar
India, New Delhi
All India Institute of Medical Sciences, new Delhi
Warren, Robin Mark
South Africa, Stellenbosch
Stellenbosch University
Bateman, E. D.
South Africa, Cape Town
University of Cape Town Lung Institute
Dheda, Keertan U.J.
South Africa, Cape Town
University of Cape Town Lung Institute
South Africa, Cape Town
University of Cape Town
United Kingdom, London
Ucl Medical School
Statistics
Citations: 100
Authors: 12
Affiliations: 5
Identifiers
Doi:
10.1371/journal.pone.0028815
e-ISSN:
19326203
Research Areas
Health System And Policy
Study Approach
Quantitative