Disch Jolande, Pedras Mariana Junqueira, Orsini Marcela, Pirmez Claude, de Oliveira Maria Cláudia, Castro Marcelo, Rabello Ana
Laboratory of Clinical Research, Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz (FIOCRUZ), Belo Horizonte, MG 30190-002, Brazil.
Diagn Microbiol Infect Dis. 2005 Mar;51(3):185-90. doi: 10.1016/j.diagmicrobio.2004.10.005.
The utility of 2 polymerase chain reaction (PCR)-based assays amplifying genus or Viannia subgenus Leishmania minicircle kDNA for the diagnostics of ML was assessed. The Viannia subgenus product was yielded after PCR from isolates of L. (Viannia) braziliensis, L. (Viannia) colombiensis, and L. (Viannia) guyanensis, whereas no product was obtained with the non-Viannia-pertaining species: L. (Leishmania) amazonensis, L. (Leishmania) donovani, and L. (Leishmania) chagasi. With both assays, 11 of 13 (86.4%) patients with confirmed ML could be identified, whereas only 2 (16.7%) of these patients were positive by microscopy. All amplified genus-specific products gave a positive signal by hybridization with a Leishmania (Viannia) subgenus-specific radioactive probe. The Viannia subgenus-specific kDNA PCR represents a sensitive and specific tool for the diagnosis of ML, remarkably improving the sensitivity of parasitological methods and offering an alternative for the radioactive-dependent assays for subgenus characterization.
评估了两种基于聚合酶链反应（PCR）的检测方法，即扩增利什曼原虫属或维阿尼亚种小环线粒体DNA（kDNA）用于黏膜皮肤利什曼病（ML）诊断的效用。PCR后，从巴西利什曼原虫（维阿尼亚种）、哥伦比亚利什曼原虫（维阿尼亚种）和圭亚那利什曼原虫（维阿尼亚种）的分离株中获得了维阿尼亚种产物，而与非维阿尼亚种相关的物种，即亚马逊利什曼原虫（利什曼原虫属）、杜氏利什曼原虫（利什曼原虫属）和恰加斯利什曼原虫（利什曼原虫属）未获得产物。通过这两种检测方法，13例确诊为ML的患者中有11例（86.4%）能够被识别出来，而通过显微镜检查，这些患者中只有2例（16.7%）呈阳性。所有扩增的属特异性产物与利什曼原虫（维阿尼亚种）属特异性放射性探针杂交后均给出阳性信号。维阿尼亚种特异性kDNA PCR是一种用于诊断ML的灵敏且特异的工具，显著提高了寄生虫学方法的灵敏度，并为亚属鉴定的放射性依赖检测提供了一种替代方法。
