Differentiation of trypanosomatid species by hybridization to selected rRNA probes.
Oligonucleotides with sequences complementary to selected regions of the Trypanosoma cruzi large sub-unit ribosomal RNA (rRNA) were used to specifically detect and quantify T. cruzi and other kinetoplastids. By selecting sequences with varying homologies with Crithida fasciculata, another kinetoplastid for which this sequence was known, probes which hybridized to T. cruzi alone or T. cruzi and T. rangeli, various Leishmania species or C. fasciculata were identified. This identification was possible even though the sequences of the large sub-unit (LSU) rRNA of T. rangeli and Leishmania are not known. None of the probes hybridized with rRNA from mouse or human cell lines, and all could quantitatively detect T. cruzi in tissue culture cells. Probing of replicate membranes with these different oligonucleotides allowed discrimination between these species. The functional application of rRNA-specific probes in diagnosis was demonstrated by identification of unknown trypanosomatids in hemocultures of wild-captured owl and squirrel monkeys using a combination of oligonucleotides. Therefore, these probes should be useful in diagnosis and identification of T. cruzi and related parasites.
Benavides
G R
GR
Department of Zoology, University of Georgia, Athens 30602.
Sullivan
J J
JJ
Steurer
F
F
McGraw
R A
RA
Tarleton
R L
RL
D000818:Animals; D001029:Aotus trivirgatus / Q000469:parasitology; D001483:Base Sequence; D002460:Cell Line; D004275:DNA, Ribosomal / Q000235:genetics; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012335:RNA, Ribosomal / Q000235:genetics*; D012453:Saimiri / Q000469:parasitology; D013045:Species Specificity; D014345:Trypanosoma / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D014349:Trypanosoma cruzi / Q000235:genetics
D004275:DNA, Ribosomal; D015345:Oligonucleotide Probes; D012335:RNA, Ribosomal