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Journal of Clinical Microbiology, September 2005, p. 4349-4356, Vol. 43, No. 9
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.9.4349-4356.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Genotyping of Madurella mycetomatis by Selective Amplification of Restriction Fragments (Amplified Fragment Length Polymorphism) and Subtype Correlation with Geographical Origin and Lesion Size

Wendy W. J. van de Sande,^1* Roy Gorkink,² Guus Simons,^2,3 Alewijn Ott,¹ Abdalla O. A. Ahmed,⁴ Henri Verbrugh,¹ and Alex van Belkum¹

Erasmus MC University Medical Center Rotterdam, Department of Medical Microbiology & Infectious Diseases, Dr. Molewaterplein 40, 3015 GD Rotterdam, The Netherlands,¹ Keygene N.V., Department of Microbial Genomics, Agro Businesspark 90, 6708 PW Wageningen, The Netherlands,² PathoFinder BV, Canisius-Wilhelmina Hospital, Weg door Jonkerbos 100, 6532 SZ Nijmegen, The Netherlands,³ Mycetoma Research Group, Institute of Endemic Diseases, and Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan⁴

Received 11 February 2005/ Returned for modification 31 March 2005/ Accepted 2 June 2005

One of the causative organisms of mycetoma is the fungus Madurella mycetomatis. Previously, extensive molecular typing studies identified Sudanese isolates of this fungus as clonal, but polymorphic genetic markers have not yet been identified. Here, we report on the selective amplification of restriction fragment (AFLP) analysis of 37 Sudanese clinical isolates of M. mycetomatis. Of 93 AFLP fragments generated, 25 were polymorphic, and 12 of these 25 polymorphic fragments were found in a large fraction of the strains. Comparative analysis resulted into a tree, composed of two main (clusters I and II) and one minor cluster (cluster III). Seventy-five percent of the strains found in cluster I originated from central Sudan, while the origin of the strains in cluster II was more heterogeneous. Furthermore, the strains found in cluster I were generally obtained from lesions larger than those from which the strains found in cluster II were obtained (chi-square test for trend, P = 0.03). Among the 12 more commonly found polymorphisms, 4 showed sequence homology with known genes. Marker A7 was homologous to an endo-1,4-beta-glucanase from Aspergillus oryzae, 97% identical markers A12 and B3 matched a hypothetical protein from Gibberella zeae, and marker B4 was homologous to casein kinase I from Danio rerio. The last marker seemed to be associated with strains originating from central Sudan (P = 0.001). This is the first report on a genotypic study where genetic markers which may be used to study pathogenicity in M. mycetomatis were obtained.

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* Corresponding author. Mailing address: Erasmus MC University Medical Center Rotterdam, Department of Medical Microbiology & Infectious Diseases, Dr. Molewaterplein 40, 3015 GD Rotterdam, The Netherlands. Phone: 00-31-10-4632177. Fax: 00-31-10-4633875. E-mail: w.vandesande{at}erasmusmc.nl.

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Journal of Clinical Microbiology, September 2005, p. 4349-4356, Vol. 43, No. 9
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.9.4349-4356.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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