Survey of ground, surface, and potable waters for the presence of Legionella species by EnviroAmpR PCR Legionella Kit, culture, and immunofluorescent staining

Lye, Dennis J.; Fout, G. Shay; Crout, Shari R.; Danielson, Richard E.; Thio, Claudia; Paszko-Kolva, C

doi:10.1016/s0043-1354(96)00252-7

Cited by 41 publications

(37 citation statements)

References 20 publications

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“…Thirty-two samples were positive using the IDS method, and 24 were positive by culture. Samples 17,18,30,33,34, and 35 were negative by IDS labeling and by culture. These samples correspond to cold domestic waters or well deep waters known to be free of Legionella.…”

Section: Resultsmentioning

confidence: 99%

Rapid Method for Enumeration of Viable Legionella pneumophila and Other Legionella spp. in Water

Delgado‐Viscogliosi

Simonart

Parent

et al. 2005

Appl Environ Microbiol

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A sensitive and specific method has been developed to enumerate viable L. pneumophila and other Legionella spp. in water by epifluorescence microscopy in a short period of time (a few hours). This method allows the quantification of L. pneumophila or other Legionella spp. as well as the discrimination between viable and nonviable Legionella. It simultaneously combines the specific detection of Legionella cells using antibodies and a bacterial viability marker (ChemChrome V6), the enumeration being achieved by epifluorescence microscopy. The performance of this immunological double-staining (IDS) method was investigated in 38 natural filterable water samples from different aquatic sources, and the viable Legionella counts were compared with those obtained by the standard culture method. The recovery rate of the IDS method is similar to, or higher than, that of the conventional culture method. Under our experimental conditions, the limit of detection of the IDS method was <176 Legionella cells per liter. The examination of several samples in duplicates for the presence of L. pneumophila and other Legionella spp. indicated that the IDS method exhibits an excellent intralaboratory reproducibility, better than that of the standard culture method. This immunological approach allows rapid measurements in emergency situations, such as monitoring the efficacy of disinfection shock treatments. Although its field of application is as yet limited to filterable waters, the double-staining method may be an interesting alternative (not equivalent) to the conventional standard culture methods for enumerating viable Legionella when rapid detection is required.

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Section: Resultsmentioning

confidence: 99%

Rapid Method for Enumeration of Viable Legionella pneumophila and Other Legionella spp. in Water

Delgado‐Viscogliosi

Simonart

Parent

et al. 2005

Appl Environ Microbiol

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“…Legionellae have been sporadically detected in groundwater (29,30,37). The increasing use of groundwater as a resource for potable water implies the need for more comprehensive data on the occurrence and distribution of legionellae in this type of environment.…”

Section: Discussionmentioning

confidence: 99%

Presence and Persistence of Legionella spp. in Groundwater

Costa¹,

Tiago²,

Costa

et al. 2005

Appl Environ Microbiol

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Groundwater samples (111) from six different boreholes located in two geographical areas were examined for the presence of legionellae over a 7-year period. The number of Legionella isolates detected was generally low. The colonization of the aquifers was not uniform, and the persistence of Legionella was independent of the hydraulic pumps and the plumbing system present in the borehole. A total of 374 isolates identified by fatty acid methyl ester analysis belonged to Legionella pneumophila, L. oakridgensis, L. sainthelensi, and L. londiniensis. In area 1, L. oakridgensis constituted the major population detected, exhibiting only one random amplified polymorphic DNA (RAPD)-PCR profile. L. sainthelensi strains were less frequently isolated and also displayed a single RAPD profile, while L. pneumophila was only sporadically detected. In contrast, L. pneumophila comprised the vast majority of the isolates in area 2 and exhibited six distinct RAPD patterns, indicating the presence of different genetic groups; three L. londiniensis RAPD types were also detected. Two of the L. pneumophila and one of the L. londiniensis RAPD types were persistent in this environment for at least 12 years. The genetic structure of L. pneumophila groundwater populations, inferred from rpoB and dotA gene sequences, was peculiar, since the majority of the isolates were allied in a discrete group different from the lineages containing most of the type and reference strains of the three subspecies of L. pneumophila. Furthermore, gene exchange events related to the dotA allele could be envisioned.

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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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Survey of ground, surface, and potable waters for the presence of Legionella species by EnviroAmpR PCR Legionella Kit, culture, and immunofluorescent staining

Cited by 41 publications

References 20 publications

Rapid Method for Enumeration of Viable Legionella pneumophila and Other Legionella spp. in Water

Rapid Method for Enumeration of Viable Legionella pneumophila and Other Legionella spp. in Water

Presence and Persistence of Legionella spp. in Groundwater

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

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