B. Oury et al., TRYPANOSOMA-CRUZI - EVALUATION OF A RAPD SYNAPOMORPHIC FRAGMENT AS A SPECIES-SPECIFIC DNA-PROBE, The Journal of parasitology, 83(1), 1997, pp. 52-57
A methodological approach is proposed to select rapidly DNA sequences
characterized by a well defined specificity and potentially interestin
g to be used as diagnostic probes or as taxonomic phylogenetic markers
. A fragment amplified from a diversified sample of Trypanosoma cruzi
stocks by the RAPD (random amplified polymorphic DNA) method with the
A8 primer, previously found to be monomorphic in all stocks, was separ
ated in 2 fragments using polyacrylamide electrophoresis. RFLP (restri
ction fragment length polymorphism) analysis of the 750-bp fragment co
mmon to all stocks revealed some sequence heterogeneity within the T.
cruzi species, whereas hybridization experiments showed a high homolog
y between fragments amplified from different T. cruzi stocks. These re
sults suggest that sequence analysis will allow the design of internal
primers to be used as probes to target specific taxonomic levels (clo
ne, family of related clones, or species) and for diagnosis.