Rapid Extraction Method of Mycobacterium ulcerans DNA from Clinical Samples of Suspected Buruli Ulcer Patients
Loading...
Date
2019-11-26
Journal Title
Journal ISSN
Volume Title
Publisher
Diagnostics
Abstract
Isothermal amplification techniques such as recombinase polymerase amplification (RPA)
and loop-mediated isothermal amplification (LAMP) for diagnosing Buruli ulcer, a necrotic skin
disease caused by Mycobacterium ulcerans, have renewed hope for the molecular diagnosis of clinically
suspected Buruli ulcer cases in endemic districts. If these techniques are applied at district-level
hospitals or clinics, they will help facilitate early case detection with prompt treatment, thereby
reducing disability and associated costs of disease management. The accuracy as well as the
application of these molecular techniques at point of need is dependent on simple and fast DNA
extraction. We have modified and tested a rapid extraction protocol for use with an already developed
recombinase polymerase amplification assay. The entire procedure from “sample in, extraction
and DNA amplification” was conducted in a mobile suitcase laboratory within 40 min. The DNA
extraction procedure was performed within 15 min, with only two manipulation/pipetting steps
needed. The diagnostic sensitivity and specificity of this extraction protocol together with M. ulcerans
RPA in comparison with standard DNA extraction with real-time PCR was 87% (n = 26) and 100%
(n = 13), respectively. We have established a simple, fast and e cient protocol for the extraction and
detection of M. ulcerans DNA in clinical samples that is adaptable to field conditions.
Description
An article published by Diagnostics
Keywords
Mycobacterium ulcerans, PCR, recombinase polymerase amplification, point-of-care
Citation
Diagnostics 2019, 9, 204; doi:10.3390/diagnostics9040204