In Thailand, asymptomatic leishmaniasis cases have continuously increased, especially among patients with HIV at risk to develop further symptoms of cutaneous and visceral leishmaniasis. Thus, earlier diagnosis using a simple, sensitive and reliable diagnostic tool is needed because populations at risk mostly reside in rural communities where only basic laboratory equipment is available for health care services. In this present study, a closed tube loop mediated isothermal amplification (LAMP) was developed using a piece of parafilm placed between the dye and LAMP reaction mixture to form semi-layer that partially secured SYBR green I from spilling during amplification. No post-amplification preparation was required and accidental spill of the dye during LAMP amplification was prevented. The result could be visually interpreted under visible and UV lights after dye spinning down. The semi-layer modification of a closed tube LAMP showed successful amplification of Leishmania DNA with clear interpretation using both color and fluorescence dyes when observing by the naked eye. The sensitivity and specificity were as high as 94.4 and 96.9%, respectively whereas detection limits were 10² parasites/mL being ten fold more sensitive than other related studies. This user-friendly inexpensive approach is affordable and suitable for empowering leishmaniasis surveillance without the need of expensive devices in all levels of hospitals, including health services, as well as fieldwork, especially in low income countries.

在泰国，无症状性利什曼病病例持续增加，尤其是在有感染艾滋病毒的患者中，有进一步发展为皮肤和内脏利什曼病症状的危险。因此，需要使用简单，灵敏和可靠的诊断工具进行早期诊断，因为处于危险中的人群大多居住在农村地区，那里只有基本的实验室设备可用于医疗服务。在本研究中，使用一块放置在染料和LAMP反应混合物之间的封口膜形成半层，从而部分封闭了SYBR green I，防止其在扩增过程中溢出，从而开发出了一个封闭的管环介导的等温扩增（LAMP）。不需要扩增后的准备，并且可以防止LAMP扩增过程中染料的意外溢出。染料旋转后，可以在可见光和紫外线下直观地解释结果。封闭管LAMP的半层修饰显示成功扩增了肉眼观察时，利什曼原虫DNA使用颜色和荧光染料具有清晰的解释。灵敏度和特异性分别高达94.4％和96.9％，而检出限为10 ^2个寄生虫/ mL，是其他相关研究灵敏度的十倍。这种用户友好的廉价方法价格合理，适用于增强利什曼病的监测，而无需在包括卫生服务以及实地调查在内的所有级别的医院中使用昂贵的设备，尤其是在低收入国家。