Point-of-Care Chlamydia trachomatis Detection Using Loop-Mediated Isothermal Amplification and Hydroxynaphthol Blue

Abstract

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.