Uliana S R, Affonso M H, Camargo E P, Floeter-Winter L M
Departamento de Parasitologia, Universidade de São Paulo, Brazil.
Exp Parasitol. 1991 Feb;72(2):157-63. doi: 10.1016/0014-4894(91)90133-h.
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.
利什曼原虫属和锥虫属基因组DNA的PvuII限制性酶切图谱分析显示出属特异性图谱。在利什曼原虫18S核糖体DNA的5'结构域中检测到一个特定的PvuII位点。合成了一个包含该PvuII区域的20聚体寡核苷酸。该序列用作探针时，在斑点试验的短曝光中，能检测到所分析的所有利什曼原虫物种的10³个完整前鞭毛体，但不与克氏锥虫或人类核酸杂交。还合成了另外两个寡核苷酸用作引物，通过聚合酶链反应扩增包含PvuII位点的18S核糖体DNA区域。所描述的探针可能有助于在临床和流行病学试验中检测利什曼原虫属。
