Use of Polymerase Chain Reaction in an Epidemiologic Investigation of Pontiac Fever

Miller, Lisa; Beebe, James L.; Butler, Jay C.; Martin, W. J.; Benson, Robert F.; Hoffman, Richard E.; Fields, Barry S.

doi:10.1093/infdis/168.3.769

Cited by 57 publications

(39 citation statements)

References 8 publications

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“…Furthermore, VBNC Legionella spp. have been shown to be capable of causing pneumonic legionellosis (14), and exposure to high concentrations of them may be an important cause of Pontiac fever (39).…”

Section: Discussionmentioning

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: Discussionmentioning

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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“…Various molecular methods have also been developed for detecting Legionella. Under certain circumstances, culture of environmental samples should be supplemented with additional tests such as polymerase chain reaction (PCR) to detect non-viable and/or viable but non-culturable legionellae (Miller et al, 1993). PCR detection may be superior to the culture and direct¯uorescent antibody methods for detecting Legionella spp.…”

Section: Detection and Disinfectionmentioning

confidence: 99%

Legionella: from environmental habitats to disease pathology, detection and control

Atlas

1999

Environmental Microbiology

210

148

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Studies on Legionella show a continuum from environment to human disease. Legionellosis is caused by Legionella species acquired from environmental sources, principally water sources such as cooling towers, where Legionella grows intracellularly in protozoa within biofilms. Aquatic biofilms, which are widespread not only in nature, but also in medical and dental devices, are ecological niches in which Legionella survives and proliferates and the ultimate sources to which outbreaks of legionellosis can be traced. Invasion and intracellular replication of L. pneumophila within protozoa in the environment play a major role in the transmission of Legionnaires' disease. Protozoa provide the habitats for the environmental survival and reproduction of Legionella species. L. pneumophila proliferates intracellularly in various species of protozoa within vacuoles studded with ribosomes, as it also does within macrophages. Growth within protozoa enhances the environmental survival capability and the pathogenicity (virulence) of Legionella. The growth requirements of Legionella, the ability of Legionella to enter a viable non‐culturable state, the association of Legionella with protozoa and the occurrence of Legionella within biofilms complicates the detection of Legionella and epidemiological investigations of legionellosis. Polymerase chain reaction (PCR) methods have been developed for the molecular detection of Legionella and used in environmental and epidemiological studies. Various physical and chemical disinfection methods have been developed to eliminate Legionella from environmental sources, but gaining control of Legionella in environmental waters, where they are protected from disinfection by growing within protozoa and biofilms, remains a challenge, and one that must be overcome in order to eliminate sporadic outbreaks of legionellosis.

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“…As we now know, these two syndromes may coexist within an exposed population ( Fig. 1) (16)(17)(18), but it is unclear whether Pontiac fever is one potential outcome in the spectrum of disease severity or whether it is due to the presence of nonviable legionellae, amoebal pathogens, and/or high levels of bacterial endotoxin (19)(20)(21)(22)(23).…”

mentioning

confidence: 99%

Current and Emerging Legionella Diagnostics for Laboratory and Outbreak Investigations

2015

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SUMMARY Legionnaires' disease (LD) is an often severe and potentially fatal form of bacterial pneumonia caused by an extensive list of Legionella species. These ubiquitous freshwater and soil inhabitants cause human respiratory disease when amplified in man-made water or cooling systems and their aerosols expose a susceptible population. Treatment of sporadic cases and rapid control of LD outbreaks benefit from swift diagnosis in concert with discriminatory bacterial typing for immediate epidemiological responses. Traditional culture and serology were instrumental in describing disease incidence early in its history; currently, diagnosis of LD relies almost solely on the urinary antigen test, which captures only the dominant species and serogroup, Legionella pneumophila serogroup 1 (Lp1). This has created a diagnostic “blind spot” for LD caused by non-Lp1 strains. This review focuses on historic, current, and emerging technologies that hold promise for increasing LD diagnostic efficiency and detection rates as part of a coherent testing regimen. The importance of cooperation between epidemiologists and laboratorians for a rapid outbreak response is also illustrated in field investigations conducted by the CDC with state and local authorities. Finally, challenges facing health care professionals, building managers, and the public health community in combating LD are highlighted, and potential solutions are discussed.

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Use of Polymerase Chain Reaction in an Epidemiologic Investigation of Pontiac Fever

Cited by 57 publications

References 8 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

Legionella: from environmental habitats to disease pathology, detection and control

Current and Emerging Legionella Diagnostics for Laboratory and Outbreak Investigations

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