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Please use this identifier to cite or link to this item: http://hdl.handle.net/123456789/11920

Title: Rapid Extraction Method of Mycobacterium ulcerans DNA from Clinical Samples of Suspected Buruli Ulcer Patients
Authors: Phillips, Richard O.
Frimpong, Michael
Ahor, Hubert Senanu
Sakyi, Samuel Asamoah
Agbavor, Bernadette
et. al
Keywords: Mycobacterium ulcerans
PCR
recombinase polymerase amplification
point-of-care
Issue Date: 26-Nov-2019
Publisher: Diagnostics
Citation: Diagnostics 2019, 9, 204; doi:10.3390/diagnostics9040204
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
URI: http://hdl.handle.net/123456789/11920
Appears in Collections:College of Health Sciences

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