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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
medicine
Operational feasibility of using loop-mediated isothermal amplification for diagnosis of pulmonary tuberculosis in microscopy centers of developing countries
Journal of Clinical Microbiology, Volume 45, No. 6, Year 2007
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Description
The characteristics of loop-mediated isothermal amplification (LAMP) make it a promising platform for the molecular detection of tuberculosis (TB) in developing countries. Here, we report on the first clinical evaluation of LAMP for the detection of pulmonary TB in microscopy centers in Peru, Bangladesh, and Tanzania to determine its operational applicability in such settings. A prototype LAMP assay with simplified manual DNA extraction was evaluated for accuracy and ease of use. The sensitivity of LAMP in smear- and culture-positive sputum specimens was 97.7% (173/177 specimens; 95% confidence interval [CI], 95.5 to 99.9%), and the sensitivity in smear-negative, culture-positive specimens was 48.8% (21/43 specimens; CI, 33.9 to 63.7%). The specificity in culture-negative samples was 99% (500/505 specimens; CI, 98.1 to 99.9%). The average hands-on time for testing six samples and two controls was 54 min, similar to that of sputum smear microscopy. The optimal amplification time was 40 min. No indeterminate results were reported, and the interreader variability was 0.4%. Despite the use of a single room without biosafety cabinets for all procedures, no DNA contamination was observed. The assay was robust, with high end-point stability and low rates of test failure. Technicians with no prior molecular experience easily performed the assay after 1 week of training, and opportunities for further simplification of the assay were identified. Copyright © 2007, American Society for Microbiology. All Rights Reserved.
Authors & Co-Authors
Boehme, Catharina C.
Switzerland, Geneva
Foundation for Innovative new Diagnostics, Switzerland
Nabeta, Pamela
Peru, Lima
Universidad Peruana Cayetano Heredia
Henostroza, German
Peru, Lima
Universidad Peruana Cayetano Heredia
Raqib, Rubhana
Bangladesh, Dhaka
International Centre for Diarrhoeal Disease Research Bangladesh
Rahim, Zeaur
Bangladesh, Dhaka
International Centre for Diarrhoeal Disease Research Bangladesh
Gerhardt, Martina
Tanzania
Mbeya Medical Research Programme
Sanga, Erica Samson
Tanzania
Mbeya Medical Research Programme
Höelscher, Michael
Germany, Munich
Ludwig-maximilians-universität München
Notomi, Tsugunori
Japan, Tokyo
Eiken Chemical Co, Ltd
Hase, Tetsu
Japan, Tokyo
Eiken Chemical Co, Ltd
Perkins, Mark D.
Switzerland, Geneva
Foundation for Innovative new Diagnostics, Switzerland
Statistics
Citations: 341
Authors: 11
Affiliations: 6
Identifiers
Doi:
10.1128/JCM.02352-06
ISSN:
00951137
Research Areas
Genetics And Genomics
Infectious Diseases
Study Locations
Tanzania