The development of a novel diagnostic PCR forMadurella mycetomatisusing a comparative genome approach

ABSTRACT Eumycetoma is a neglected tropical disease characterized by large tumorous lesions. It is most commonly caused by the fungus Madurella mycetomatis which accounts for more than 70% of cases in central Africa. Currently, identification of the causative agent can only be reliably performed by a species-specific PCR. However, we recently demonstrated that our M. mycetomatis specific PCR can cross-react with Madurella pseudomycetomatis. We therefore used a comparative genome approach to develop a new M. mycetomatis specific PCR for species identification. For this we compared the published M. mycetomatis genome to genomes of other organisms in BLASTCLUST to identify unique M. mycetomatis predicted protein coding sequences. Based on 16 of these unique sequences, PCR primers were developed. The specificity of these primers was further evaluated in other eumycetoma causing agents including the Madurella sibling species. Out of the 16 tested sequences, only one was unique for M. mycetomatis and this should be used as a novel diagnostic marker for M. mycetomatis.