Premium
Chip‐based duplex real‐time PCR for water quality monitoring concerning Legionella pneumophila and Legionella spp.
Author(s) -
Reuter Cornelia,
Hentschel Stefanie,
Breitenstein Antje,
Heinrich Eileen,
Aehlig Oliver,
Henkel Thomas,
Csáki Andrea,
Fritzsche Wolfgang
Publication year - 2021
Publication title -
water and environment journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.437
H-Index - 37
eISSN - 1747-6593
pISSN - 1747-6585
DOI - 10.1111/wej.12635
Subject(s) - legionella , legionella pneumophila , real time polymerase chain reaction , polymerase chain reaction , microbiology and biotechnology , biology , bacteria , genetics , gene
Based on biomolecular methods, rapid and selective identification of human pathogenic water organisms becomes an important issue. Legionella spp., are pathogenic water bacteria with worldwide significance. Prevalent detection methods for these microorganisms are time and/or cost intensive. We describe a detection setup and relating DNA assay. A miniaturized real‐time polymerase chain reaction (real‐time PCR) for direct on‐line discrimination of Legionella pneumophila and Legionella spp. was established and integrated into a real‐time PCR‐chip‐system. The PCR‐chip device combines a temperature controlling unit and a fluorescence intensity measurement. It was designed to achieve rapid amplification, using an approach of real‐time fluorescence read out with the intercalating dye EvaGreen® and melting curve analysis, without requiring multiple probes. The presented results exhibit reproducibility and good sensitivity, showing that the setup is suitable for robust, rapid and cost‐efficient detection and monitoring of a variety of Legionella spp.in urban water samples.