Detection and classification of Trypanosoma cruzi by DNA hybridization with nonradioactive probes.
Total or kinetoplast DNA (kDNA) from 72 isolates and clones of Trypanosoma cruzi as well as from nine related trypanosomatids were analyzed by dot hybridization using nonradioactive kDNA or cloned minicircle fragments as probes. Biotinylated-kDNA probes generated by nick-translation proved reliable for distinguishing Zymodeme 1 and Zymodeme 2bol of T. cruzi parasites. In contrast, digoxigenin-labeled kDNA obtained by random-priming did not distinguish among T. cruzi isolates but did distinguish among New World leishmanias. Cloned minicircle fragments labeled with digoxigenin gave the same results as digoxigenin-labeled kDNA, except for a 10-fold decrease in sensitivity. Digoxigenin-labeled DNA probes proved useful in unambiguously detecting T. cruzi from different geographic regions of America. However, T. rangeli and T. cruzi marinkellei were not distinguished by these probes.
Solari
A
A
Departamento de Bioquimica, Facultad de Medicina Norte, Universidad de Chile, Santiago.
Venegas
J
J
Gonzalez
E
E
Vasquez
C
C
D000818:Animals; D001483:Base Sequence; D014355:Chagas Disease / Q000175:diagnosis; D003001:Cloning, Molecular; D015342:DNA Probes; D004270:DNA, Circular; D018105:DNA, Kinetoplast; D016054:DNA, Protozoan / Q000032:analysis*; D004076:Digoxigenin; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D015341:Nucleic Acid Probes; D014349:Trypanosoma cruzi / Q000145:classification / Q000302:isolation & purification*
D015342:DNA Probes; D004270:DNA, Circular; D018105:DNA, Kinetoplast; D016054:DNA, Protozoan; D015341:Nucleic Acid Probes; D004076:Digoxigenin