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Article Dans Une Revue Epidemiology Année : 2006

Hospital Network Water Monitoring. Detection of Legionella Pneumophila by a Miniarray-PCR Assay

Romain R. Bertrand
Benoit Roig

Résumé

Legionella are gram-negative bacteria found in soils and freshwater. Legionella genus has been established in 1979 after pneumonia outbreak in American Legion members. Species pneumophila causes 90% of legionellosis and serogroup 1 is detected in approximately 80% of the cases. Legionella are facultative intracellular development bacteria, growths in environment within amoeba and human phagocytic cells. Multiplication within amoeba allows it to survive in environment. It causes respiratory diseases in human when a susceptible host inhales contaminated aerosolized water or aspirates water. Approximately 10000–40000 people in the United States develop Legionnaires’ disease annually. In France, about 1200 cases a year are registered with 20 to 30% mortality. Bacterium transmission is possible in equipments at risk such as hot water tanks and storages, showers, cooling towers, spa rooms, humidifiers and condensate tanks, respiratory nebulizers, face spritzers, ice generators, fountain or bottle water coolers. Human contamination is possible when sensitive persons for their health status and/or their treatment are strongly and/or frequently exposured to contaminated water aerosols. The standard method for Legionella detection (ISO 11–731) is the laboratory culture, with 250 CFU/liter of sensitivity. The main drawbacks of the method are its slowness and culture difficulties of bacteria. In spite of a great collection of clinical and environmental detection techniques, few methods combine rapidity with specificity and sensitivity. Qualitative or real-time quantitative PCR (Polymerase Chain Reaction) allows a rapid, specific and sensitive detection. Moreover, the use of DNA microarray technology enhances the detection capabilities, especially in term of sensitivity. The method presented here uses DNA amplification and labeling by PCR of a dnaJ gene fragment followed by hybridization on probe modified membrane and its visualization by a biotin colorimetric system. About 200 hospital water samples are analysed by this method and the reference method (ISO 11–731). Results obtained by the two methods are compared by statistic considerations. The miniarray system allows to obtain results in the day for species diagnostic. An internal control allows to ovoid false-negative results, to validate template DNA preparation, PCR and hybridization steps. This rapid detection method should allow proceeding to auto-surveillance system of Legionella presence on hospital network water. So if positive results a corrective action could be started right away. It should be adapted to alert system and should reply to requirements of infectious risk control.
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hal-01663979 , version 1 (14-12-2017)

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  • HAL Id : hal-01663979 , version 1

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Romain R. Bertrand, Benoit Roig. Hospital Network Water Monitoring. Detection of Legionella Pneumophila by a Miniarray-PCR Assay. Epidemiology, 2006, 17 (6), pp.S392. ⟨hal-01663979⟩
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