Identification of Trypanosoma cruzi zymodemes by kinetoplast DNA probes.
Analysis of zymograms of extracts of Trypanosoma cruzi isolated from different hosts in Argentina allowed characterization of 12 zymodemes or "isozymic strains," only six of which were found in human patients. Two of these six zymodemes (Z1 and Z12) were widely distributed and found in more than 80% of human patients. These two "major natural clones" differed significantly in pathogenic activity. Because the groupings obtained by studying enzymes and kinetoplast DNA (kDNA) were similar, it is possible to identify the zymodeme by analyzing kDNA. A 290-bp fragment was amplified by PCR using primers for the sequences flanking the hypervariable regions of kDNA minicircles. Labeled probes for this fragment, prepared from Z1 and Z12 reference stocks, hybridized specifically with PCR-amplified kDNA from parasite stocks, allowing identification of zymodemes.
Montamat
E E
EE
Cátedra de Química Biológica, Facultad de Ciencias Médicas, Universidad Nacional de Córdoba, Argentina.
Durand
S
S
Bocco
J L
JL
De Luca D'Oro
G M
GM
Blanco
A
A
D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D014355:Chagas Disease / Q000469:parasitology*; D015342:DNA Probes; D018105:DNA, Kinetoplast / Q000032:analysis*; D005260:Female; D006801:Humans; D007527:Isoenzymes / Q000032:analysis* / Q000235:genetics; D008297:Male; D008875:Middle Aged; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods; D014349:Trypanosoma cruzi / Q000145:classification / Q000201:enzymology* / Q000235:genetics*
D015342:DNA Probes; D018105:DNA, Kinetoplast; D007527:Isoenzymes