Than Linh Quyen
Technical University of Denmark
Denmark
Title: Selection of DNA dyes for rapid and sensitive real-time Loop-Mediated Isothermal Amplification
Biography
Biography: Than Linh Quyen
Abstract
Loop-mediated isothermal amplification (LAMP) has received great interest and is wildly used for point of care (POC) diagnostics based nucleic acid detection of pathogens. For example, DNA dye is an indispensable component in real-time LAMP reaction. Several DNA dyes work well for real-time PCR, but they have inhibition effect in LAMP reaction. Hence, the selection of dyes is urgent need for development of rapid, accurate, sensitive real-time LAMP based quantification and detection of pathogens. In this study, we have investigated twenty four DNA dyes for rapid real-time LAMP detection of Salmonella enteritidis based on inhibition effects. The real-time LAMP reactions were performed with 24 dyes at various concentrations ranged from 0.5 µM to 10 µM. Among the 24 dyes tested, SYTO dye group (except SYTO 60 and SYTO 62), Miami dye group (except Miami Red), and Boxto were the best since non-inhibition effect on the real-time LAMP reaction was observed. Application of one of the best dyes, the SYTO 9, in the real-time LAMP reaction, a detection limit of 3.86 copies of hilA gene for Salmonella spp. within 30 min at 65 °C was archived. The identification of the best dyes for rapid real-time LAMP detection will contribute important information for development of POC real-time LAMP systems for robust, sensitive and rapid online or at site detection of the pathogens.