title: Madurella mycetomatis strains from mycetoma lesions in Sudanese patients are clonal.
abstract: Molecular diversity among clinical isolates of Madurella mycetomatis, the prime fungal agent of human mycetoma in Sudan, could possibly explain the diverse clinical presentations of this severely debilitating infectious disease. In addition, culture-independent DNA-mediated typing tests need to be developed for this organism, since M. mycetomatis DNA, but not the organism itself, can be identified in soil, the material from which infections are thought to originate. A collection of 38 different clinical M. mycetomatis isolates was characterized by large-scale random amplification of polymorphic DNA using 20 different primer species. These analyses, involving at least 2,600 annealing sites, showed a complete lack of DNA fingerprint variation among the various isolates. From the resulting homogeneous DNA fingerprints, seven fragments were cloned and sequenced, and novel, species-specific PCR restriction fragment length polymorphism (RFLP) tests were designed. The seven PCR RFLP tests were successfully performed on the 38 different M. mycetomatis strains. However, again all M. mycetomatis DNA patterns obtained appeared to be identical, whereas patterns produced using DNAs from other fungal species were clearly discriminatory. These results suggest that there is little genetic variation among clinically relevant M. mycetomatis strains from Sudan. The data tentatively imply that different manifestations of mycetoma are due to differences in host susceptibility rather than differential virulence of the causative agent.
authors.affiliation: Mycetoma Research Group, Institute of Endemic Diseases, University of Khartoum, Khartoum, Sudan.
authors.forename: Abdalla
authors.initials: A
authors.lastname: Ahmed
authors.forename: Wendy
authors.initials: W
authors.lastname: van de Sande
authors.forename: Henri
authors.initials: H
authors.lastname: Verbrugh
authors.forename: Ahmed
authors.initials: A
authors.lastname: Fahal
authors.forename: Alex
authors.initials: A
authors.lastname: van Belkum
mesh_terms: D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis; D006801:Humans; D020461:Madurella / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D008271:Mycetoma / Q000382:microbiology*; D016533:Mycological Typing Techniques; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D019105:Random Amplified Polymorphic DNA Technique; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013397:Sudan
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal