Simple, Rapid Mycobacterium ulcerans Disease Diagnosis from Clinical Samples by Fluorescence of Mycolactone on Thin Layer Chromatography

Abstract

Introduction Mycobacterium ulcerans infection, known as Buruli ulcer, is a disease of the skin and subcutaneous tissues which is an important but neglected tropical disease with its major impact in rural parts of West and Central Africa where facilities for diagnosis and management are poorly developed. We evaluated fluorescent thin layer chromatography (f-TLC) for detection of mycolactone in the laboratory using samples from patients with Buruli ulcer and patients with similar lesions that gave a negative result on PCR for the IS2404 repeat sequence of M. ulcerans Methodology/Principal findingsMycolactone and DNA extracts from fine needle aspiration (FNA), swabs and biopsy specimen were used to determine the sensitivity and specificity of f-TLC when compared with PCR for the IS2404. For 71 IS2404 PCR positive and 28 PCR negative samples the sensitivity was 73.2% and specificity of 85.7% for f-TLC. The sensitivity was similar for swabs (73%), FNAs (75%) and biopsies (70%).ConclusionsWe have shown that mycolactone can be detected from M. ulcerans infected skin tissue by f-TLC technique. The technique is simple, easy to perform and read with minimal costs. In this study it was undertaken by a member of the group from each endemic country. It is a potentially implementable tool at the district level after evaluation in larger field studies.

Global Epidemiology of Buruli Ulcer, 2010–2017, and Analysis of 2014 WHO Programmatic Targets

“…A point-of-care diagnostic tool is needed and would greatly improve confirmation of Buruli ulcer cases in the field. Currently, simpler methods such as loop-mediated isothermal amplification assay and fluorescent thin layer chromatography are being tested in some treatment centers in Africa (37).…”

Integrated Control and Management of Neglected Tropical Skin Diseases

“…Fluorescent thin layer chromatography (fTLC) is a simple and low-cost technique that can be used for detection of mycolactone in skin swabs from BU lesions at a peripheral hospital laboratory using a small bench analyser;[30] however, this test is still in the development stage. The Dual Path Platform (DPP) yaws rapid test kit, which is based on simultaneous detection of antibodies to treponemal and nontreponemal antigens, allows for serological diagnosis of yaws in the field.…”

TranslocatingMycobacterium ulcerans: an experimental model

“…This experimental result corroborates the fact that M. ulcerans has never been detected in a tissue or organ at a distance from its inoculation point, which also suggests that the cytotoxic activity of mycolactones is local and not systemic, without clarifying the role of mycobacterial inhibition and mycolactone inhibition themselves. Interestingly, it is possible to determine mycolactones in Buruli ulcer lesions, but not in patients’ blood [16]. It should be noted that translocation property is shared with mycobacteria of the Mycobacterium tuberculosis complex, in which experimental translocation has been shown for Mycobacterium canettii [17] and M. tuberculosis [18], there are no experimental data or clinical observations to our knowledge for mycobacteria of the Mycobacterium leprae complex.…”