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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
immunology and microbiology
Pyrazinamide susceptibility testing of Mycobacterium tuberculosis by high resolution melt analysis
Tuberculosis, Volume 94, No. 1, Year 2014
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Description
Summary Pyrazinamide (PZA) plays the important role in shortening the tuberculosis treatment period and in treating MDR-TB. Phenotypic PZA susceptibility methods are limited because they require specialized acidified media, which increases costs and complexity. In this study we developed a genotypic high resolution melt (HRM) analysis technique to detect pncA mutations associated with PZA resistant Mycobacterium tuberculosis. Seven overlapping primer pairs were designed to cover the entire pncA gene and upstream regions. Each gene segment was individually amplified by real-time PCR followed by HRM analysis. The assay was evaluated on 98 clinical M. tuberculosis isolates (41 PZA susceptible by MGIT method, 55 PZA resistant, 2 undetermined). HRM was 94% concordant to full-length sequencing results, with most discrepancies attributable to mixed populations per HRM or transversions. Sequencing and HRM yielded 82% and 84% concordance, respectively, to phenotypic PZA susceptibilities by MGIT, with most discrepancies attributable to isolates with wild-type pncA but phenotypic PZA resistance. This HRM technique is a simple and high-throughput method for screening clinical M. tuberculosis samples for PZA resistance. © 2013 Elsevier Ltd. All rights reserved.
Authors & Co-Authors
Pholwat, Suporn
United States, Charlottesville
University of Virginia
Stroup, Suzanne E.
United States, Charlottesville
University of Virginia
Gratz, Jean C.
United States, Charlottesville
University of Virginia
Trangan, Varittha
Thailand, Bangkok
Siriraj Hospital
Foongladda, Suporn
Thailand, Bangkok
Siriraj Hospital
Kumburu, Happiness Houka
Tanzania, Moshi
Kilimanjaro Clinical Research Institute
Juma, Saumu Pazia
Tanzania, Moshi
Kilimanjaro Clinical Research Institute
Kibiki, Gibson S.
Tanzania, Moshi
Kilimanjaro Clinical Research Institute
Houpt, Eric R.
United States, Charlottesville
University of Virginia
Statistics
Citations: 23
Authors: 9
Affiliations: 3
Identifiers
Doi:
10.1016/j.tube.2013.10.006
ISSN:
14729792
e-ISSN:
1873281X
Research Areas
Genetics And Genomics
Infectious Diseases