Detection of Legionellae in Hospital Water Samples by Quantitative Real-Time LightCycler PCR

Wellinghausen, Nele; Frost, Cathrin; Marre, R.

doi:10.1128/aem.67.9.3985-3993.2001

Cited by 187 publications

(184 citation statements)

References 23 publications

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“…Moreover, several authors have described the isolation of Legionella spp from showers, cooling towers and boilers (26,33,52), which is in agreement with the findings of the present study. In contrast with previous findings reported in literature (4,51), Legionella spp were not detected in dental units water.…”

Section: Discussionsupporting

confidence: 83%

Detection of Legionella pneumophila in water and biofilm samples by culture and molecular methods from man-made systems in São Paulo - Brazil

Carvalho

Foronda

Pellizari³

2007

Braz. J. Microbiol.

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Legionella pneumophila is a pathogenic bacteria associated to aquatic habitat of natural and artificial environments. Clinical cases of legionellosis have been reported in Brazil but there is a lack of information about the incidence and concentration of this bacterium in environmental sources. Thus, the present study was designed to evaluate the occurrence of legionellae in São Paulo city, Brazil, using different methods of detection and identification. Sixty-seven water and biofilm samples from natural reservoirs and man-made systems were collected and analyzed for the presence of Legionella spp by culturing onto a selective medium, coculture in axenic free-living amoebae and direct fluorescent antibody (DFA) assay. Results showed that freshwater of reservoirs did not contain legionellae, Legionella pneumophila was isolated from man-made systems, with predominance of Legionella pneumophila serogroup 1 strains. Although there was no statistical difference among the proposed detection methods, the plate culture method yielded a higher number of L. pneumophila positive samples, followed by amoebic coculture procedure and direct fluorescent antibody assay. Results of PCR and sequencing reactions revealed that application of macrophage infectivity potentiator gene as a molecular marker was an important tool for the identification of environmental isolates of L. pneumophila. The agreement among the three detection methods-when all methods yielded similar results-and the prevalence of a single Legionella species in the sampled man-made systems could suggest that the occurrence of this bacterium had been influenced by the higher concentration of metallic ions dissociated in water of those systems than in natural reservoirs. Thus, the results of this study revealed that the water of man-made systems in Sao Paulo may serve as a reservoir for L. pneumophila and other microorganism, including free-living protozoans.

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Section: Discussionsupporting

confidence: 83%

Detection of Legionella pneumophila in water and biofilm samples by culture and molecular methods from man-made systems in São Paulo - Brazil

Carvalho

Foronda

Pellizari³

2007

Braz. J. Microbiol.

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“…L. pneumophila primers were Lp-mip-PT69 (5Ј-GCA TTG GTG CCG ATT TGG-3Ј) and Lp-mip-PT70 (5Ј-GYT TTG CCA TCA AAT CTT TCT (52). Standards for the Legionella and L.p.Ϫmip assay were generated with DNA extracted from a plate scrape of L. pneumophila subsp.…”

Section: Its Gene Sequencingmentioning

confidence: 99%

Opportunistic pathogens enriched in showerhead biofilms

Feazel

Baumgartner²,

Peterson³

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

497

364

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The environments we humans encounter daily are sources of exposure to diverse microbial communities, some of potential concern to human health. In this study, we used culture-independent technology to investigate the microbial composition of biofilms inside showerheads as ecological assemblages in the human indoor environment. Showers are an important interface for human interaction with microbes through inhalation of aerosols, and showerhead waters have been implicated in disease. Although opportunistic pathogens commonly are cultured from shower facilities, there is little knowledge of either their prevalence or the nature of other microorganisms that may be delivered during shower usage. To determine the composition of showerhead biofilms and waters, we analyzed rRNA gene sequences from 45 showerhead sites around the United States. We find that variable and complex, but specific, microbial assemblages occur inside showerheads. Particularly striking was the finding that sequences representative of non-tuberculous mycobacteria (NTM) and other opportunistic human pathogens are enriched to high levels in many showerhead biofilms, >100-fold above background water contents. We conclude that showerheads may present a significant potential exposure to aerosolized microbes, including documented opportunistic pathogens. The health risk associated with showerhead microbiota needs investigation in persons with compromised immune or pulmonary systems.bioaerosols ͉ Mycobacterium avium complex ͉ Non-tuberculous mycobacteria ͉ public health ͉ rRNA metagenomics

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“…A number of PCR-based assays have been developed for the detection and quantification of Legionella in water mainly using the 5S and 16S rRNA genes and the macrophage infectivity potentiator (mip) gene of L. pneumophila [23][24][25][26][27][28][29][30][31]. However, these assays lack the ability to discriminate between living and dead (non-infectious) Legionella cells.…”

Section: Introductionmentioning

confidence: 99%

Identification of Legionella spp. in Environmental Water Samples by ScanVIT-Legionella™ Method in Spain

Grúas

Alvarez²,

Lara

et al. 2013

Indian J Microbiol

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Rapid and more sensitive methods for the detection and quantification of viable Legionella cells have been developed. In this paper, a comparative analysis of environmental water samples using the ScanVIT-Legionella TM method and the traditional ''gold standard'' method of culturing is realised indicating the usefulness of the ScanVIT method. The ScanVIT-Legionella TM method was performed on environmental water samples from different locations of Huesca region (Spain). Legionella micro-colonies should appear green colour and Legionella pneumophila micro-colonies appear red. Twenty-one environmental water samples analysed by standard culture plus five control samples (Two sterile water samples with Legionella as positive controls and three sterile water samples as negative controls). All of them were used to apply ScanVIT-Legionella TM method. From of 21 environmental samples eleven were positive, six negative with both methods and four samples were negative for culture method and positive for ScanVIT-Legionella TM method. The positive control samples were positive and the negative were negative for both methods. A comparative analysis of the results obtained with two methods showed a strong positive determination coefficient (R 2 = 0.99753). The results demonstrate the usefulness of the ScanVITLegionella TM method as a rapid diagnostic tool in order to provide a diagnosis as quick as possible. ScanVITLegionella TM method offers a series of advantages such as quickly diagnosis, higher sensitivity and the possibility to identify Legionella spp. and L. pneumophila simultaneously.

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Detection of Legionellae in Hospital Water Samples by Quantitative Real-Time LightCycler PCR

Cited by 187 publications

References 23 publications

Detection of Legionella pneumophila in water and biofilm samples by culture and molecular methods from man-made systems in São Paulo - Brazil

Detection of Legionella pneumophila in water and biofilm samples by culture and molecular methods from man-made systems in São Paulo - Brazil

Opportunistic pathogens enriched in showerhead biofilms

Identification of Legionella spp. in Environmental Water Samples by ScanVIT-Legionella™ Method in Spain

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