Abstract
Mycolactones, macrolide cytotoxins, are key virulence factors of Mycobacterium ulcerans, the etiological agent of the chronic necrotizing skin disease Buruli ulcer. There is urgent need for a simple point-of-care laboratory test for Buruli ulcer and mycolactone represents a promising target for the development of an immunological assay. However, for a long time, all efforts to generate mycolactone-specific antibodies have failed. By using a protein conjugate of a truncated non-toxic synthetic mycolactone derivative, we recently described generation of a set of mycolactone-specific monoclonal antibodies. Using the first mycolactone-specific monoclonal antibodies that we have described before, we were able to develop an antigen competition assay that detects mycolactones. By the systematic selection of a capturing antibody and a reporter molecule, and the optimization of assay conditions, we developed an ELISA that detects common natural variants of mycolactone with a limit of detection in the low nanomolar range. The mycolactone-specific ELISA described here will be a very useful tool for research on the biology of this macrolide toxin. After conversion into a simple point-of-care test format, the competition assay may have great potential as laboratory assay for both the diagnosis of Buruli ulcer and for the monitoring of treatment efficacy.
Highlights
IntroductionMycobacterium ulcerans is the etiological agent of the chronic necrotizing skin disease Buruli ulcer (BU) that primarily affects children in West and Central Africa [1]
Using the first mycolactone-specific monoclonal antibodies that we have described previously, we have optimized here an antigen competition ELISA that detects common natural variants of mycolactone at a low nanomolar scale
Mycobacterium ulcerans is the etiological agent of the chronic necrotizing skin disease Buruli ulcer (BU) that primarily affects children in West and Central Africa [1]
Summary
Mycobacterium ulcerans is the etiological agent of the chronic necrotizing skin disease Buruli ulcer (BU) that primarily affects children in West and Central Africa [1]. Genomic analyses have shown that M. ulcerans has emerged from a common ancestor with the fish pathogen Mycobacterium marinum [2, 3] by acquisition of a virulence plasmid carrying genes that encode polyketide-modifying enzymes and the giant polyketide synthases responsible for the synthesis of the lipid toxin mycolactone [4]. Mycolactone plays a key role in the chronic necrotizing pathogenesis of BU and, in addition, analgesic and immunosuppressive effects are attributed to the toxin [6]. At an air/buffer interface, mycolactone has been shown to have surfactant properties with an apparent surface saturation concentration of 1 μM [8]
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