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Lett Appl Microbiol. 2005;41(6):470-5.

Detection of Legionella pneumophila in water samples by species-specific real-time and nested PCR assays.

Author information

1
ARPA, Regional Agency for Environmental Protection and Health Prevention, Emilia Romagna Region, Bologna, Italy. lfiume@bo.arpa.emr.it

Erratum in

  • Lett Appl Microbiol. 2006 Mar;42(3):304. Bucca Sabattini, MA [corrected to Bucci Sabattini, MA].

Abstract

AIMS:

Legionella pneumophila is a contaminant of man-made water systems, including potable water, cooling towers, water systems of large buildings, etc. It is the most common causative agent of legionellosis, a respiratory infection, which may give rise to restricted outbreaks. To survey environmental water samples from hospitals and private habitations in Bologna, we developed a species-specific nested and a TaqMan real-time PCR for the detection of L. pneumophila. We compared the two assays and both to cultural isolation.

METHODS AND RESULTS:

The targeted gene was macrophage infectivity potentiator (mip), conserved in L. pneumophila, and divergent in other legionellae. One assay was based on a nested PCR and the other on a TaqMan real-time PCR protocol. Their sensitivities were 14 % or 5% higher than that of cultural isolation respectively. The detection limits were 1-2 genome equivalents per 50-microl reaction. Specificity was assessed using DNA from nine target and 20 nontarget organisms.

CONCLUSIONS:

When applied to water samples, both assays detected L. pneumophila at 80% or higher frequency.

SIGNIFICANCE AND IMPACT OF THE STUDY:

The species-specific molecular diagnosis of L. pneumophila by means of nested PCR does not require a specific instrumentation, exhibits a high sensitivity, and is advantageous over the cultural isolation and real-time PCR detection. It allows to quickly monitor water samples for the risk assessment of environmental contaminations.

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