Genomix Molecular Diagnostics (P) Ltd., 5-36/207 Prasanthnagar, Kukatpally, Hyderabad 500 072, India.
Exp Parasitol. 2013 Jul;134(3):333-40. doi: 10.1016/j.exppara.2013.03.031. Epub 2013 Apr 4.
We attempted to improve the loop-mediated isothermal amplification (LAMP) method for malaria diagnosis by using a simple DNA extraction procedure, and a portable device performing both the amplification and detection of LAMP in one platform. Additionally, the device served as a heating block for the DNA preparation. We refer this method as LAMP-Tube scanner, and evaluated using 209 microscopically positive malaria samples and compared them to RDTs and LAMP-Thermocycler. Two most common human infecting Plasmodium species were detected. The LAMP-Tube scanner method is found to be simple and allowed real-time detection of DNA amplification. The time to amplification varied but was closely less than 60 min. Sensitivity and specificity of LAMP-Tube scanner in detecting Plasmodium falciparum were 95% and 93.3%, compared to microscopy and 98.3% and 100% respectively, compared to standard LAMP-Thermocycler. In addition, it showed a detection limit of 10 and 40 copies of the parasitemia for Plasmodium vivax and P. falciparum. Accordingly, in comparison to the results obtained by microscopy, the LAMP-Tube scanner had a less divergence in sensitivity and specificity, and yielded results similar to those of LAMP-Thermocycler. This method has the great potential as a field usable molecular tool for the diagnosis of malaria and is an alternative to conventional PCR-based diagnostic methods for field use.
我们尝试通过使用简单的 DNA 提取程序和一个能够在同一平台上同时执行扩增和检测 LAMP 的便携式设备来改进用于疟疾诊断的环介导等温扩增(LAMP)方法。此外,该设备还可用作 DNA 制备的加热块。我们将这种方法称为 LAMP-Tube 扫描仪,并使用 209 份显微镜阳性疟疾样本进行评估,并将其与 RDT 和 LAMP-Thermocycler 进行比较。该方法能够检测到两种最常见的感染人类的疟原虫。LAMP-Tube 扫描仪方法简单,允许实时检测 DNA 扩增。扩增时间有所不同,但均紧密少于 60 分钟。LAMP-Tube 扫描仪检测恶性疟原虫的灵敏度和特异性分别为 95%和 93.3%,与显微镜相比,分别为 98.3%和 100%,与标准 LAMP-Thermocycler 相比。此外,它显示出对间日疟原虫和恶性疟原虫的寄生虫血症分别为 10 和 40 拷贝的检测限。因此,与显微镜获得的结果相比,LAMP-Tube 扫描仪在灵敏度和特异性方面的差异较小,并且产生的结果与 LAMP-Thermocycler 相似。该方法具有作为现场可用的疟疾分子诊断工具的巨大潜力,是现场使用的基于常规 PCR 的诊断方法的替代方法。