Evaluation of different DNA extraction methods and loop-mediated isothermal amplification primers for the detection of Mycobacterium ulcerans in clinical specimens
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Date
2021
Journal Title
Journal ISSN
Volume Title
Publisher
BMC Infectious Diseases
Abstract
Background: Early diagnosis and treatment of Buruli ulcer is critical in order to avoid the debilitating effects of the
disease. In this regard, the development of new diagnostic and point of care tools is encouraged. The loopmediated
isothermal amplification for the detection of Mycobacterium ulcerans represents one of the new tools
with a good potential of being developed into a point of care test. There is however the need to standardize the
assays, reduce sample preparation times, improve the detection/visualization system and optimize them for highthroughput
screening, adaptable to low resourced laboratories.
Methods: In this study, we assessed two DNA extraction protocols (modified Boom and EasyNAT methods), three
previously published LAMP primer sets (BURULI, MU 2404 and BU-LAMP), and compared the sensitivity and
specificity of LAMP assays on three DNA amplification platforms.
Results: Our results show that Buruli ulcer diagnosis using primers targeting IS2404 for the LAMP method is
sensitive (73.75–91.49%), depending on the DNA extraction method used. Even though the modified Boom DNA
extraction method provided the best results, its instrumentation requirement prevent it from being field applicable.
The EasyNAT method on the other hand is simpler and may represent the best method for DNA extraction in less
resourced settings.
Conclusions: For further work on the development and use of LAMP tests for Buruli diagnosis, it is recommended
that the BURULI sets of primers be used, as these yielded the best results in terms of sensitivity (87.50–91.49%) and
specificity (89.23–100%), depending on the DNA extraction methods used.
Description
Research Article
Keywords
Loop mediated isothermal amplification, DNA extraction, Sensitivity, Specificity