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Rapid detection of viable Legionella pneumophila in tap water by a qPCR and RT‐PCR‐based method
Author(s) -
Boss R.,
Baumgartner A.,
Kroos S.,
Blattner M.,
Fretz R.,
Moor D.
Publication year - 2018
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.13932
Subject(s) - legionella pneumophila , legionella , legionnaires' disease , tap water , microbiology and biotechnology , biology , real time polymerase chain reaction , 16s ribosomal rna , polymerase chain reaction , bacteria , environmental science , gene , environmental engineering , biochemistry , genetics
Aims A molecular method for a rapid detection of viable Legionella pneumophila of all serogroups in tap water samples was developed as an alternative to the reference method (ISO). Legionellae are responsible for Legionnaires’ disease, a severe pneumonia in humans with high lethality. Methods and Results The developed method is based on a nutritional stimulation and detection of an increase in precursor 16S rRNA as an indicator for viability. For quantification, DNA was detected by qPCR. This method was compared to the ISO method using water samples obtained from public sports facilities in Switzerland. The sensitivity and specificity were 91 and 97%, respectively, when testing samples for compliance with a microbiological criterion of 1000 cell equivalents per l. Conclusion The new method is sensitive and specific for Leg. pneumophila and allows results to be obtained within 8 h upon arrival, compared to one week or more by the ISO method. Significance and Impact of the Study The method represents a useful tool for a rapid detection of viable Leg. pneumophila of all serogroups in water by molecular biology. It can be used as an alternative to the ISO method for official water analysis for legionellae and particularly when a short test time is required.