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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
medicine
Rapid detection of Mycobacterium tuberculosis and rifampin resistance by use of on-demand, near-patient technology
Journal of Clinical Microbiology, Volume 48, No. 1, Year 2010
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Description
Current nucleic acid amplification methods to detect Mycobacterium tuberculosis are complex, labor-intensive, and technically challenging. We developed and performed the first analysis of the Cepheid Gene Xpert System's MTB/RIF assay, an integrated hands-free sputum-processing and real-time PCR system with rapid on-demand, near-patient technology, to simultaneously detect M. tuberculosis and rifampin resistance. Analytic tests of M. tuberculosis DNA demonstrated a limit of detection (LOD) of 4.5 genomes per reaction. Studies using sputum spiked with known numbers of M. tuberculosis CFU predicted a clinical LOD of 131 CFU/ml. Killing studies showed that the assay's buffer decreased M. tuberculosis viability by at least 8 logs, substantially reducing biohazards. Tests of 23 different commonly occurring rifampin resistance mutations demonstrated that all 23 (100%) would be identified as rifampin resistant. An analysis of 20 nontuberculosis mycobacteria species confirmed high assay specificity. A small clinical validation study of 107 clinical sputum samples from suspected tuberculosis cases in Vietnam detected 29/29 (100%) smear-positive culture-positive cases and 33/39 (84.6%) or 38/53 (71.7%) smear-negative culture-positive cases, as determined by growth on solid medium or on both solid and liquid media, respectively. M. tuberculosis was not detected in 25/25 (100%) of the culture-negative samples. A study of 64 smear-positive culture-positive sputa from retreatment tuberculosis cases in Uganda detected 63/64 (98.4%) culture-positive cases and 9/9 (100%) cases of rifampin resistance. Rifampin resistance was excluded in 54/55 (98.2%) susceptible cases. Specificity rose to 100% after correcting for a conventional susceptibility test error. In conclusion, this highly sensitive and simple-to-use system can detect M. tuberculosis directly from sputum in less than 2 h. Copyright © 2010, American Society for Microbiology. All Rights Reserved.
Available Materials
https://efashare.b-cdn.net/share/pmc/articles/PMC2812290/bin/supp_48_1_229__index.html
https://efashare.b-cdn.net/share/pmc/articles/PMC2812290/bin/supp_48_1_229__genexpert_supplermentary_methods_danicapaper_JCM_sub_2_fin.doc
Authors & Co-Authors
Helb, Danica A.
United States, Newark
Rutgers new Jersey Medical School
United States, Berkeley
University of California, Berkeley
Jones, Martin
United States, Sunnyvale
Cepheid
Story, Elizabeth
United States, Newark
Rutgers new Jersey Medical School
Boehme, Catharina C.
Switzerland, Geneva
Foundation for Innovative new Diagnostics, Switzerland
Wallace, Ellen
United States, Sunnyvale
Cepheid
Ho, Ken
United States, Sunnyvale
Cepheid
Kop, Joann
United States, Sunnyvale
Cepheid
Owens, Michelle R.
United States, Sunnyvale
Cepheid
Rodgers, Richard
United States, Sunnyvale
Cepheid
Banada, Padmapriya P.
United States, Newark
Rutgers new Jersey Medical School
Safi, Hassan
United States, Newark
Rutgers new Jersey Medical School
Blakemore, Robert
United States, Newark
Rutgers new Jersey Medical School
Ngoc Lan, N. T.
Viet Nam, Ho Chi Minh City
Pham Ngoc Thach Hospital
Jones-López, Edward C.
United States, Newark
Rutgers new Jersey Medical School
Levi, Michael
United States, New York
Montefiore Medical Center
Burday, Michele
United States, Newark
Rutgers new Jersey Medical School
Ayakaka, Irene
Uganda, Kampala
Makerere University
Mugerwa, Roy D.
Uganda, Kampala
School of Medicine, Makerere University College of Health Sciences
McMillan, Bill
United States, Sunnyvale
Cepheid
United States, Cupertino
Profusa Inc.
Winn-Deen, Emily
United States, Sunnyvale
Cepheid
Christel, Lee
United States, Sunnyvale
Cepheid
Dailey, Peter J.
United States, Sunnyvale
Cepheid
Perkins, Mark D.
Switzerland, Geneva
Foundation for Innovative new Diagnostics, Switzerland
Persing, David H.
United States, Sunnyvale
Cepheid
Alland, David
United States, Newark
Rutgers new Jersey Medical School
Statistics
Citations: 952
Authors: 25
Affiliations: 9
Identifiers
Doi:
10.1128/JCM.01463-09
ISSN:
00951137
Research Areas
Genetics And Genomics
Health System And Policy
Study Locations
Uganda