A modified AFLP for Trypanosoma congolense isolate characterisation.
The amplified fragment length polymorphism (AFLP) technique is a reliable and powerful DNA fingerprint tool for genetic characterisation and analysis. In this paper, we described a modified AFLP with high resolution for Trypanosoma congolense using one enzyme and agarose or Elchrom gel electrophoresis. Eleven allopatric and fourteen sympatric isolates of T. congolense savannah were used to assess the resolution of the method and its ability to characterise T. congolense isolates. Two enzymes (Eco RI or Bgl II) and corresponding non-selective and selective primers were used to identify the most appropriate combination. Patterns generated by Bgl II enzyme and a single selective primer A, C, G or T produced clear profiles. Each of the four selective primers produced different profiles for all the 25 T. congolense isolates. Due to the reduction in the number of bands, profiles could be analysed using agarose or Elchrom gels. Although comparison of a great number of samples could benefit from software help, this technique did not require flurochrome detection methods. The results of the present study demonstrated that this modified AFLP makes the characterisation of T. congolense easier while maintaining high resolution.
Masumu
J
J
Institute of Tropical Medicine, Animal health Department, Nationalestraat 155, B-2000 Antwerp, Belgium.
Geysen
D
D
Vansnick
E
E
Geerts
S
S
Van den Bossche
P
P
D000818:Animals; D017931:DNA Primers / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics* / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004586:Electrophoresis / Q000379:methods; D004587:Electrophoresis, Agar Gel / Q000379:methods; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D015203:Reproducibility of Results; D014348:Trypanosoma congolense / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification
D017931:DNA Primers; D016054:DNA, Protozoan; D015252:Deoxyribonucleases, Type II Site-Specific; C018492:GCCNNNNNGGC-specific type II deoxyribonucleases