Somboonna Naraporn, Choopara Ilada
Department of Microbiology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
Methods Mol Biol. 2019;2042:11-17. doi: 10.1007/978-1-4939-9694-0_2.
Loop-mediated isothermal amplification (LAMP) is a new nucleic acid amplification technique that provides high sensitivity and specificity compatible to conventional polymerase chain reaction (PCR) and simplicity, without the requirement of an expensive thermal cycler. Additionally, LAMP coupled hydroxynaphthol blue (HNB) allowed for visual color detection by the naked eye. Here we describe the methodology of Chlamydia trachomatis detection of vaginal swab from crude DNA lysis to LAMP-HNB reaction. The result can be read by the naked eye through color change from violet (negative) to sky blue (positive). The result can be confirmed by UV spectra and agarose gel electrophoresis. This assay uses all nonhazardous chemical reagents and is hence safe to the users, and requires little specialist training or knowledge.
环介导等温扩增技术(LAMP)是一种新的核酸扩增技术,具有与传统聚合酶链反应(PCR)相当的高灵敏度和特异性,且操作简单,无需昂贵的热循环仪。此外,LAMP与羟基萘酚蓝(HNB)联用可实现肉眼可视的颜色检测。在此,我们描述了从粗DNA裂解到LAMP-HNB反应检测阴道拭子中沙眼衣原体的方法。结果可通过颜色从紫色(阴性)变为天蓝色(阳性)由肉眼读取。该结果可通过紫外光谱和琼脂糖凝胶电泳进行确认。此检测方法使用的均为无危险的化学试剂,因此对使用者安全,且几乎不需要专业培训或知识。