Loop-mediated amplification as promising on-site detection approach for Legionella pneumophila and Legionella spp

Appl Microbiol Biotechnol. 2020 Jan;104(1):405-415. doi: 10.1007/s00253-019-10286-3. Epub 2019 Dec 12.

Abstract

Recently Legionella pneumophila is the main causative waterborne organism of severe respiratory infections. Additionally, other Legionella species are documented as human pathogens. In our work, we describe a rapid detection method which combines two advantages for sensitive and specific detection of the genus Legionella: the fast isothermal amplification method "Loop-mediated isothermal AMPlification" (LAMP), and a colorimetric detection method using the metal indicator hydroxynaphtol blue (HBN) which allows to determine an optical signal with a simple readout (with the naked eye). Moreover, we present two approaches for minimizing the assay volume using a stationary microchip LAMP and droplet digital-based LAMP (ddLAMP) as promising highly sensitive setups.

Keywords: Droplet digital–based amplification; Loop-mediated isothermal amplification; Naked eye.

MeSH terms

  • Colorimetry
  • DNA Primers / genetics
  • Legionella / isolation & purification*
  • Legionella pneumophila / isolation & purification*
  • Naphthalenesulfonates / metabolism
  • Nucleic Acid Amplification Techniques / methods*
  • Sensitivity and Specificity
  • Temperature

Substances

  • DNA Primers
  • Naphthalenesulfonates
  • trisodium 3-hydroxy-4-((2Z)-2-(2-oxo-4-sulfonatonaphthalen-1-ylidene)hydrazinyl)naphthalene-2,7-disulfonate