A quick and cost effective method for the diagnosis of <it>Mycobacterium ulcerans </it>infection

• Main Authors: de Souza Dziedzom K, Quaye Charles, Mosi Lydia, Addo Phyllis, Boakye Daniel A
• Format: Article
• Language: English
• Published: BMC 2012-01-01
• Series: BMC Infectious Diseases
• Subjects: Buruli ulcer; <it>Mycobacterium ulcerans</it>; Diagnosis; Loop mediated isothermal amplification
• Online Access: http://www.biomedcentral.com/1471-2334/12/8

<p>Abstract</p> <p>Background</p> <p>Buruli ulcer (BU), a neglected tropical skin disease caused by <it>Mycobacterium ulcerans</it>, has been reported in over 30 countries worldwide and is highly endemic in rural West and Central Africa. The mode of transmission remains unknown and treatment is the only alternative to disease control. Early and effective treatment to prevent the morbid effects of the disease depends on early diagnosis; however, current diagnosis based on clinical presentation and microscopy has to be confirmed by PCR and other tests in reference laboratories. As such confirmed BU diagnosis is either late, inefficient, time consuming or very expensive, and there is the need for an early diagnosis tool at point of care facilities. In this paper we report on a simple, quick and inexpensive diagnostic test that could be used at point of care facilities, in resource-poor settings.</p> <p>Methods</p> <p>The methodology employed is based on the loop mediated isothermal amplification (LAMP) technique. Four sets of Primers, targeting the mycolactone encoding plasmid genome sequence of <it>M. ulcerans </it>were designed. The BU-LAMP assay was developed and tested on five <it>M. ulcerans </it>strains from patients in Ghana and two American Type Culture Control (ATCC) reference isolates; Ghana #970321 (D19F9) and Benin #990826 (D27D14). We also tested the assay on other closely related, mycolactone-producing mycobacterial strains; <it>M. marinum </it>1218, <it>M. marinum </it>DL240490, <it>M. liflandii </it>and <it>M. pseudoshotsii</it>, as well as experimentally infected laboratory animal and clinical samples.</p> <p>Results</p> <p>The results revealed a high specificity of the BU-LAMP assay for selectively detecting <it>M. ulcerans</it>. Compared to the conventional IS-2404 PCR, the new assay is cheaper and simpler and ten times more sensitive. Test results can be obtained within 1 hour.</p> <p>Conclusions</p> <p>This study indicates that the BU-LAMP assay could be suitable for early disease diagnosis and application in low-resource health facilities.</p>