Detection of
            <i>Legionella pneumophila</i>
            Using a Real-Time PCR Hybridization Assay

Ballard, Anna; Fry, Norman K.; Chan, LC; Surman, Susanne; Lee, J. V.; Harrison, Timothy G.; Towner, K. J.

doi:10.1128/jcm.38.11.4215-4218.2000

Cited by 96 publications

(33 citation statements)

References 14 publications

(12 reference statements)

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“…However, specific bacterial species are more frequently the focus for real-time PCR assays, especially when long culture times can be replaced by rapid and specific gene detection. Leptospira genospecies, Mycobacterium and Propionibacterium spp., Chlamydia spp., Legionella pneumophila and Listeria monocytogenes have all been detected and in some cases quantitated with the use of real-time PCR assays [41,[234][235][236][237][238][239][240][241][242][243][244][245][246].…”

Section: Bacteriamentioning

confidence: 99%

Real-time PCR in the microbiology laboratory

Mackay

2004

Clinical Microbiology and Infection

643

338

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Use of PCR in the field of molecular diagnostics has increased to the point where it is now accepted as the standard method for detecting nucleic acids from a number of sample and microbial types. However, conventional PCR was already an essential tool in the research laboratory. Real-time PCR has catalysed wider acceptance of PCR because it is more rapid, sensitive and reproducible, while the risk of carryover contamination is minimised. There is an increasing number of chemistries which are used to detect PCR products as they accumulate within a closed reaction vessel during real-time PCR. These include the non-specific DNA-binding fluorophores and the specific, fluorophore-labelled oligonucleotide probes, some of which will be discussed in detail. It is not only the technology that has changed with the introduction of real-time PCR. Accompanying changes have occurred in the traditional terminology of PCR, and these changes will be highlighted as they occur. Factors that have restricted the development of multiplex real-time PCR, as well as the role of real-time PCR in the quantitation and genotyping of the microbial causes of infectious disease, will also be discussed. Because the amplification hardware and the fluorogenic detection chemistries have evolved rapidly, this review aims to update the scientist on the current state of the art. Additionally, the advantages, limitations and general background of real-time PCR technology will be reviewed in the context of the microbiology laboratory.

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Section: Bacteriamentioning

confidence: 99%

Real-time PCR in the microbiology laboratory

Mackay

2004

Clinical Microbiology and Infection

643

338

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“…Several qPCR assays targeting Leg. pneumophila have been developed during the last few years (Ballard et al 2000;Wellinghausen et al 2001;Levi et al 2003;Joly et al 2006;Behets et al 2007). The use of qPCR in the direct enumeration of Leg.…”

Section: Introductionmentioning

confidence: 99%

“…The use of qPCR in the direct enumeration of Leg. pneumophila was evaluated in sample of natural water, hospital water and cooling tower water (Ballard et al 2000;Wellinghausen et al 2001;Levi et al 2003;Joly et al 2006). Moreover, some authors reported that the interpretation of qPCR could be influenced by the type of water samples (e.g.…”

Section: Introductionmentioning

confidence: 99%

Evaluation ofLegionella pneumophilacontamination in Italian hotel water systems by quantitative real-time PCR and culture methods

Bonetta

Ferretti

Balocco

et al. 2010

Journal of Applied Microbiology

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Aims: This study was designed to define the extent of water contamination by Legionella pneumophila of certain Italian hotels and to compare quantitative real‐time PCR with the conventional culture method. Methods and Results: Nineteen Italian hotels of different sizes were investigated. In each hotel three hot water samples (boiler, room showers, recycling) and one cold water sample (inlet) were collected. Physico‐chemical parameters were also analysed. Legionella pneumophila was detected in 42% and 74% of the hotels investigated by the culture method and by real‐time PCR, respectively. In 21% of samples analysed by the culture method, a concentration of >104 CFU l−1 was found, and Leg. pneumophila serogroup 1 was isolated from 10·5% of the hotels. The presence of Leg. pneumophila was significantly influenced by water sample temperature, while no association with water hardness or residual‐free chlorine was found. Conclusions: This study showed a high percentage of buildings colonized by Leg. pneumophila. Moreover, real‐time PCR proved to be sensitive enough to detect lower levels of contamination than the culture method. Significance and Impact of the Study: This study indicates that the Italian hotels represent a possible source of risk for Legionnaires’ disease and confirms the sensitivity of the molecular method. To our knowledge, this is the first report to demonstrate Legionella contamination in Italian hotels using real‐time PCR and culture methods.

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“…On these occasions, a rapid detection of contamination is needed to minimize the risk of case appearance and ⁄ or to check the effectiveness of any corrective measures. Although the culture technique represents the gold standard, it takes up to 10 days before results are available and has a limited sensitivity, and inter-laboratory variations have been reported in diagnostic field, but not on water samples (Boulanger and Edelstein 1995;Ballard et al 2000;Tronel and Hartemann 2009). Thus, a more rapid, simple and reproducible technique for legionellae detection in environmental samples may be of special interest for routine laboratory applications.…”

mentioning

confidence: 99%