Department of Medical Laboratory Sciences, Faculty of Health Sciences, Hokkaido University, Kita-12 Nishi-5, Kita-ku, Sapporo, Hokkaido, 060-0812, Japan.
Division of Bioresources, Hokkaido University Research Center for Zoonosis Control, Kita 20 Nishi 10, Kita-ku, Sapporo, Hokkaido, 001-0020, Japan; German Nepal Tuberculosis Project, Kalimati, Kathmandu, Nepal; Healthy Nepal, Balkhu, Kathmandu, Nepal.
Tuberculosis (Edinb). 2019 Jul;117:1-6. doi: 10.1016/j.tube.2019.05.004. Epub 2019 May 21.
The purpose of this study was to develop a simple visual methyl green (MeG) based dry loop-mediated isothermal amplification (LAMP) method for early detection of Mycobacterium tuberculosis (MTB) from clinical samples. We identified MeG as an indicator of a positive LAMP reaction, where a positive reaction gave a blue-green color while a negative reaction was colorless. The MeG MTB-LAMP system was further simplified by drying all reagents for ease of use, and was then validated for its ability to diagnose TB directly using Nepalese clinical samples. We evaluated the dry MeG MTB-LAMP with 69 new TB suspected samples from patients that did not have a confirmed history of TB treatment and found the sensitivity in culture positive samples as 92.8% (13/14) and specificity in culture negative samples as 96.3% (53/55). Our LAMP system has the potential to be a point of care test for early diagnosis of active TB in developing countries.
本研究旨在开发一种简单的基于甲基绿（MeG）的可视化干环介导等温扩增（LAMP）方法，用于从临床样本中早期检测结核分枝杆菌（MTB）。我们发现 MeG 是 LAMP 反应阳性的指示剂，阳性反应呈蓝绿色，而阴性反应则无色。通过将所有试剂干燥以简化 MeG-MTB-LAMP 系统，使其更便于使用，然后使用尼泊尔临床样本验证其直接诊断结核病的能力。我们评估了干 MeG-MTB-LAMP 在 69 例新的疑似结核病患者样本中的应用，这些患者没有明确的结核病治疗史，在培养阳性样本中的灵敏度为 92.8%（13/14），在培养阴性样本中的特异性为 96.3%（53/55）。我们的 LAMP 系统有可能成为发展中国家早期诊断活动性结核病的即时检测方法。
