Application of flow cytometry to detection and characterization of Legionella spp

Tyndall, R. L.; Hand, Russell E.; Mann, Riti; Evans, C. Lovatt; Jernigan, Robert W.

doi:10.1128/aem.49.4.852-857.1985

Cited by 61 publications

(19 citation statements)

References 11 publications

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“…From a taxonomic point of view there are not, as far as we know, any studies concerning the taxonomic discrimination of individual components of natural bacteria populations. The only research on the condition of protocols allowing differentiation of taxonomic units (type, species) was carried out on mixtures of pure strains, notably for diagnostic of pathogenic bacteria [14,24,36,50,58]. The determination of the proportions of the bases of DNA (AT/GC), by means of specific fluorochromes (eg AT, Hoechst 33258 and DAPI; GC, chromomycin A3 and mithramycin), would allow the discrimination of certain bacterial species [53].…”

Section: Type Of Study Stain(s) or Method(s) And Reference(s) For Bacmentioning

confidence: 99%

“…However, two organisms that show similar AT/GC ratios are not necessarily identical since these proportions do not take into account the linear arrangement of nucleotides in DNA [25]. The use of monoclonal or polyclonal antibodies [58,62] and nucleic fluorescent probes [2] is a priori certainly more promising. An example of detection of a strain of Salmonella typhimurium identified by means of indirect immunofluorescent staining following the methods of Desmonts et al [12] is shown in figure 2.…”

Section: Type Of Study Stain(s) or Method(s) And Reference(s) For Bacmentioning

confidence: 99%

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Recent applications of flow cytometry in aquatic microbial ecology

Troussellier¹,

Courties²,

Vaquer³

1993

Biology of the Cell

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Microorganisms (unicellular algae, bacteria) constitute fundamental compartments of aquatic ecosystems because of their high concentrations and activities. The evaluation and understanding of their behavior and role raise different problems for which traditional methodologies are often inadequate, whether they refer to global or classical microscopic analyses. Flow cytometry (FCM) has been recently used to study microorganisms in aquatic environments. Although this technology is still applied on a limited scale in our field, a large number of works has been done showing that FCM seems to be a promising tool for aquatic microbial ecology. This paper summarizes, from the literature produced during the last decade and with original data obtained in our laboratory, the main questions related to the cell identification, the evaluation of cell viability, biomasses and productions and the measurements of bacterial and phytoplanktonic activities. The representatives of sampling and observation scales is also discussed within the framework of the FCM measurements.

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Section: Type Of Study Stain(s) or Method(s) And Reference(s) For Bacmentioning

confidence: 99%

Section: Type Of Study Stain(s) or Method(s) And Reference(s) For Bacmentioning

confidence: 99%

Recent applications of flow cytometry in aquatic microbial ecology

Troussellier¹,

Courties²,

Vaquer³

1993

Biology of the Cell

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“…Flow cytometry (FCM) was already used 30 years ago for Lp detection. These first approaches, however, had very high LODs of 10 4 to 10 6 cells per litre (Tyndall et al, 1985;Lebaron et al, 1998). More recently, FCM-based approaches reached detection limits as low as 500 cells per litre, but high background signals, not well discriminated cell clusters, high counts of false-positives and lacking information about physiological state are some of the drawbacks of these methods (Füchslin et al, 2010;Taguri et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Rapid detection of total and viableLegionella pneumophilain tap water by immunomagnetic separation, double fluorescent staining and flow cytometry

Keserue

Baumgärtner

Felleisen

et al. 2012

Microbial Biotechnology

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SummaryWe developed a rapid detection method for Legionella pneumophila (Lp) by filtration, immunomagnetic separation, double fluorescent staining, and flow cytometry (IMS‐FCM method). The method requires 120 min and can discriminate ‘viable’ and ‘membrane‐damaged’ cells. The recovery is over 85% of spiked Lp SG 1 cells in 1 l of tap water and detection limits are around 50 and 15 cells per litre for total and viable Lp, respectively. The method was compared using water samples from house installations in a blind study with three environmental laboratories performing the ISO 11731 plating method. In 53% of the water samples from different taps and showers significantly higher concentrations of Lp were detected by flow cytometry. No correlation to the plate culture method was found. Since also ‘viable but not culturable’ (VNBC) cells are detected by our method, this result was expected. The IMS‐FCM method is limited by the specificity of the used antibodies; in the presented case they target Lp serogroups 1–12. This and the fact that no Lp‐containing amoebae are detected may explain why in 21% of all samples higher counts were observed using the plate culture method. Though the IMS‐FCM method is not yet fit to completely displace the established plating method (ISO 11731) for routine Lp monitoring, it has major advantages to plating and can quickly provide important insights into the ecology of this pathogen in water distribution systems.

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“…Unlike the study of lymphocytes, little work has been done on combining FCM and monoclonal antibodies for the detection of bacteria (Tyndall et al 1985;Phillips and Martin 1988;Nelson et al 1991) and even less using the two for detection in foods (Donnelly and Baigent 1986).…”

Section: Introductionmentioning

confidence: 99%

Detection of low levels of specific Salmonella species by fluorescent antibodies and flow cytometry

McClelland

Pinder

1994

Journal of Applied Bacteriology

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The use of fluorescently-labelled monoclonal antibodies, with detection by multi-parameter flow cytometry, was investigated for the rapid detection of salmonellas in pure cultures. Accurate detection of specific Salmonella serotypes was demonstrated down to levels of below 10(4) cells ml-1 (within 30 min) and 1 cell ml-1 (after 6 h non-selective pre-enrichment). This level of sensitivity was attained even in the presence of high levels of other bacterial species that would otherwise have interfered with the results. With combinations of different antibodies, each with a unique fluorescent label, simultaneous analysis for two species was possible.

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Application of flow cytometry to detection and characterization of Legionella spp

Cited by 61 publications

References 11 publications

Recent applications of flow cytometry in aquatic microbial ecology

Recent applications of flow cytometry in aquatic microbial ecology

Rapid detection of total and viableLegionella pneumophilain tap water by immunomagnetic separation, double fluorescent staining and flow cytometry

Detection of low levels of specific Salmonella species by fluorescent antibodies and flow cytometry

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