Toubanaki Dimitra K, Athanasiou Evita, Karagouni Evdokia
Laboratory of Cellular Immunology, Department of Microbiology, Hellenic Pasteur Institute, 127 Vas. Sofias Ave., 11521 Athens, Greece.
J Microbiol Methods. 2016 Aug;127:51-58. doi: 10.1016/j.mimet.2016.05.027. Epub 2016 May 30.
Leishmaniasis is a disease, caused by Leishmania parasites, which infect humans and animals, posing a major social and economic burden worldwide. The need for accurate and sensitive disease diagnosis led to the widespread adoption of PCR amplification. Detection of the amplification products (i.e. gel electrophoresis) require time-consuming protocols performed by trained personnel, with high cost. Aim of the present study was the simplification of PCR product detection, using a nucleic acid lateral flow, combined with functionalized gold nanoparticles. Amplification reactions targeting kinetoplastid DNA of Leishmania spp were performed on canine blood samples and a positive signal was formed as a red test zone. The visual detection was completed in 20min. Extensive optimization enabled the detection of 100fmol of target DNA. Clinical samples of infected dog blood were analyzed with high specificity. Overall, the proposed lateral flow biosensor can be considered an appealing alternative platform for Leishmania-specific amplification products detection with low cost and attractive simplicity.
利什曼病是一种由利什曼原虫寄生虫引起的疾病，可感染人类和动物，在全球范围内造成重大的社会和经济负担。对准确、灵敏的疾病诊断的需求导致了聚合酶链反应（PCR）扩增技术的广泛应用。检测扩增产物（即凝胶电泳）需要由经过培训的人员执行耗时的操作流程，成本高昂。本研究的目的是使用核酸侧向流动技术并结合功能化金纳米颗粒来简化PCR产物检测。针对利什曼原虫动基体DNA的扩增反应在犬类血液样本上进行，阳性信号以红色测试区的形式出现。视觉检测在20分钟内完成。经过广泛优化，能够检测到100飞摩尔的目标DNA。对感染犬类血液的临床样本进行分析时具有很高的特异性。总体而言，所提出的侧向流动生物传感器可被视为一种有吸引力的替代平台，用于以低成本和极具吸引力的简单方式检测利什曼原虫特异性扩增产物。
