Leishmania: genus identification based on a specific sequence of the 18S ribosomal RNA sequence.
The analysis of PvuII restriction patterns of Leishmania spp. and Trypanosoma spp. genomic DNA showed genus distinctive profiles. A specific PvuII site was detected in the 5' domain of 18S ribosomal DNA of Leishmania. A 20-mer oligonucleotide encompassing this PvuII region was synthesized. This sequence, when utilized as probe, on short exposures of dot tests, detected 10(3) whole promastigotes of all Leishmania species analyzed but did not hybridize with T. cruzi or human nucleic acids. Two other oligonucleotides were synthesized to be used as primers for amplification through polymerase chain reaction of the 18S ribosomal DNA region containing the PvuII site. The probes described may be useful for the detection of Leishmania spp. under clinical and epidemiological trials.
Uliana
S R
SR
Departamento de Parasitologia, Universidade de São Paulo, Brazil.
Affonso
M H
MH
Camargo
E P
EP
Floeter-Winter
L M
LM
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D016054:DNA, Protozoan / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis*; D015252:Deoxyribonucleases, Type II Site-Specific; D007891:Leishmania / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D016053:RNA, Protozoan / Q000032:analysis; D012337:RNA, Ribosomal, 18S / Q000032:analysis*; D015183:Restriction Mapping
D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D015345:Oligonucleotide Probes; D016053:RNA, Protozoan; D012337:RNA, Ribosomal, 18S; C025630:CAGCTG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific