Cambridge Journals Online

Molecular variation of Trypanosoma brucei subspecies as revealed by AFLP fingerprinting

E. E. C.  AGBO  a1 c1 , P. A. O.  MAJIWA  a3 , H. J. H. M.  CLAASSEN  a2 and M. F. W.  TE PAS  a1 a1 Division of Animal Sciences, Section for Animal Genomics, Institute for Animal Science and Health, ID-Lelystad, Edelhertweg 15, 8200 AB Lelystad, The Netherlands a2 Department of Immunology, Erasmus University Rotterdam, Dr Molewaterplein 50, 3015 GE Rotterdam, The Netherlands a3 International Livestock Research Institute, P.O. Box 30709, Nairobi, Kenya

Article author query agbo e     majiwa p     claassen h     te pas m

Abstract

Genetic analysis of Trypanosoma spp. depends on the detection of variation between strains. We have used the amplified fragment length polymorphism (AFLP) technique to develop a convenient and reliable method for genetic characterization of Trypanosome (sub)species. AFLP accesses multiple independent sites within the genome and would allow a better definition of the relatedness of different Trypanosome (sub)species. Nine isolates (3 from each T. brucei subspecies) were tested with 40 AFLP primer combinations to identify the most appropriate pairs of restriction endonucleases and selective primers. Primers based on the recognition sequences of EcoRI and BglII were chosen and used to analyse 31 T. brucei isolates. Similarity levels calculated with the Pearson correlation coefficient ranged from 15 to 98%, and clusters were determined using the unweighted pair-group method using arithmetic averages (UPGMA). At the intraspecific level, AFLP fingerprints were grouped by numerical analysis in 2 main clusters, allowing a clear separation of T. b. gambiense (cluster I) from T. b. brucei and T. b. rhodesiense isolates (cluster II). Interspecies evaluation of this customized approach produced heterogeneous AFLP patterns, with unique genetic markers, except for T. evansi and T. equiperdum, which showed identical patterns and clustered together.