Ramos A, Maslov D A, Fernandes O, Campbell D A, Simpson L
Howard Hughes Medical Institute, University of California, Los Angeles, 90095-1662, USA.
Exp Parasitol. 1996 Apr;82(3):242-50. doi: 10.1006/expr.1996.0031.
A single pair of PCR primers within a conserved region of the mini-exon repeat was used to amplify the repeats from 10 species of pathogenic Leishmania belonging to four major clinical groups and also from three species of Trypanosoma. Oligonucleotide hybridization probes for the detection and identification of the PCR-amplified repeats were constructed from alignments of mini-exon intron and intergenic sequences. The probes generated from mini-exon intergenic regions of the L. (V.) braziliensis, L. (L.) donovani, and L. (L.) mexicana species hybridized specifically to their cognate groups without discriminating between the species within the groups. The probes for L. (L.) major and L. (L.) aethiopica were species-specific, while the L. (L.) tropica probe also hybridized with the L. (L.) aethiopica mini-exon repeat. The mini-exon intron-derived probes for T. cruzi, T. rangeli, and T. brucei were species-specific. This method involving the detection of specific PCR-amplified products produced using a single primer set represents a novel sensitive and specific assay for multiple trypanosomatid species and groups.
在小外显子重复序列的保守区域内使用一对PCR引物，从属于四个主要临床组的10种致病性利什曼原虫以及三种锥虫中扩增重复序列。根据小外显子内含子和基因间序列的比对构建了用于检测和鉴定PCR扩增重复序列的寡核苷酸杂交探针。从巴西利什曼原虫、杜氏利什曼原虫和墨西哥利什曼原虫物种的小外显子基因间区域产生的探针特异性地与其同源组杂交，而不区分组内的物种。硕大利什曼原虫和埃塞俄比亚利什曼原虫的探针具有物种特异性，而热带利什曼原虫的探针也与埃塞俄比亚利什曼原虫的小外显子重复序列杂交。克氏锥虫、兰氏锥虫和布氏锥虫的小外显子内含子衍生探针具有物种特异性。这种涉及检测使用单一引物组产生的特异性PCR扩增产物的方法代表了一种针对多种锥虫物种和组的新型灵敏且特异的检测方法。
