利什曼病患者各种临床样本中利什曼原虫的简化分子检测
Simplified molecular detection of Leishmania parasites in various clinical samples from patients with leishmaniasis.
作者信息
Mugasa Claire M, Laurent Thierry, Schoone Gerard J, Basiye Frank L, Saad Alfarazdeg A, El Safi Sayda, Kager Piet A, Schallig Henk Dfh
机构信息
Koninklijk Instituut voor de Tropen (KIT)/Royal Tropical Institute, KIT Biomedical Research, Amsterdam, the Netherlands.
出版信息
Parasit Vectors. 2010 Mar 2;3(1):13. doi: 10.1186/1756-3305-3-13.
BACKGROUND
Molecular methods to detect Leishmania parasites are considered specific and sensitive, but often not applied in endemic areas of developing countries due to technical complexity. In the present study isothermal, nucleic acid sequence based amplification (NASBA) was coupled to oligochromatography (OC) to develop a simplified detection method for the diagnosis of leishmaniasis. NASBA-OC, detecting Leishmania RNA, was evaluated using clinical samples from visceral leishmaniasis patients from East Africa (n = 30) and cutaneous leishmaniasis from South America (n = 70) and appropriate control samples.
RESULTS
Analytical sensitivity was 10 parasites/ml of spiked blood, and 1 parasite/ml of culture. Diagnostic sensitivity of NASBA-OC was 93.3% (95% CI: 76.5%-98.8%) and specificity was 100% (95% CI: 91.1%-100%) on blood samples, while sensitivity and specificity on skin biopsy samples was 98.6% (95% CI: 91.2%-99.9%) and 100% (95% CI: 46.3%-100%), respectively.
CONCLUSION
The NASBA-OC format brings implementation of molecular diagnosis of leishmaniasis in resource poor countries one step closer.
背景
检测利什曼原虫寄生虫的分子方法被认为具有特异性和敏感性，但由于技术复杂性，在发展中国家的流行地区往往未被应用。在本研究中，基于核酸序列的等温扩增技术（NASBA）与寡色层分析技术（OC）相结合，开发了一种用于利什曼病诊断的简化检测方法。使用来自东非内脏利什曼病患者（n = 30）和南美皮肤利什曼病患者（n = 70）的临床样本以及适当的对照样本，对检测利什曼原虫RNA的NASBA-OC方法进行了评估。
结果
加标血液中分析灵敏度为每毫升10个寄生虫，培养物中为每毫升1个寄生虫。NASBA-OC对血液样本的诊断灵敏度为93.3%（95%置信区间：76.5%-98.8%），特异性为100%（95%置信区间：91.1%-100%），而对皮肤活检样本的灵敏度和特异性分别为98.6%（95%置信区间：91.2%-99.9%）和100%（95%置信区间：46.3%-100%）。
结论
NASBA-OC形式使资源匮乏国家实施利什曼病的分子诊断又迈进了一步。
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