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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
medicine
Next-generation ion torrent sequencing of drug resistance mutations in Mycobacterium tuberculosis strains
Journal of Clinical Microbiology, Volume 50, No. 12, Year 2012
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Description
A novel protocol for full-length Mycobacterium tuberculosis gene analysis of first- and second-line drug resistance was developed using the Ion Torrent Personal Genome Machine (PGM). Five genes - rpoB (rifampin), katG (isoniazid), pncA (pyrazinamide), gyrA (ofloxacin/fluoroquinolone), and rrs (aminoglycosides) - were amplified and sequenced, and results were compared to those obtained by genotypic Hain line probe assay (LPA) and phenotypic Bactec MGIT 960 analysis using 26 geographically diverse South African clinical isolates collected between July and November 2011. Ion Torrent sequencing exhibited 100% (26/26) concordance to phenotypic resistance obtained by MGIT 960 culture and genotypic rpoB and katG results by LPA. In several rifampin-resistant isolates, Ion Torrent sequencing revealed uncommon substitutions (H526R and D516G) that did not have a defined mutation by LPA. Importantly, previously uncharacterized mutations in rpoB (V194I), rrs (G878A), and pncA (Q122Stop) genes were observed. Ion Torrent sequencing may facilitate tracking and monitoring geographically diverse multidrug-resistant and extensively drug-resistant strains and could potentially be integrated into selected regional and reference settings throughout Africa, India, and China. Copyright © 2012, American Society for Microbiology. All Rights Reserved.
Authors & Co-Authors
Daum, Luke T.
United States, San Antonio
Longhorn Vaccines and Diagnostics
Rodriguez, J. D.
United States, San Antonio
Longhorn Vaccines and Diagnostics
Worthy, S. A.
United States, San Antonio
Longhorn Vaccines and Diagnostics
Ismail, Nazir Ahmed
South Africa, Pretoria
University of Pretoria
South Africa, Sandringham
National Institute for Communicable Disease
Omar, Shaheed Vally
South Africa, Pretoria
University of Pretoria
South Africa, Sandringham
National Institute for Communicable Disease
Dreyer, Andries William
South Africa, Pretoria
University of Pretoria
Fourie, P. B.
South Africa, Pretoria
University of Pretoria
Hoosen, Anwar Ahmed
South Africa, Pretoria
University of Pretoria
Chambers, J. P.
United States, San Antonio
Longhorn Vaccines and Diagnostics
United States, San Antonio
The University of Texas at San Antonio
Fischer, G. W.
United States, San Antonio
Longhorn Vaccines and Diagnostics
Statistics
Citations: 77
Authors: 10
Affiliations: 4
Identifiers
Doi:
10.1128/JCM.01893-12
ISSN:
00951137
e-ISSN:
1098660X
Research Areas
Cancer
Genetics And Genomics