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Med Princ Pract. 2008;17(1):49-55.

Legionella in clinical specimens and hospital water supply facilities: molecular detection and genotyping of the isolates.

Author information

1
Department of Applied Medical Sciences, College of Health Sciences, The Public Authority for Applied Education and Training, Kuwait University, Kuwait. qasemj@paaet.edu.kw

Abstract

OBJECTIVE:

To evaluate genus- and species-specific polymerase chain reactions (PCRs) for the detection of the genus Legionella and the species Legionella pneumophila in clinical specimens and hospital water supplies, and to establish a simple and reproducible random amplification of polymorphic DNA (RAPD)-PCR technique for genotyping of Legionella.

MATERIALS AND METHODS:

A total of 70 respiratory tract specimens(bronchoalveolar lavage: n = 46; endotracheal secretions: n = 9; sputum: n = 15) from patients with atypical pneumonia, and 283 environmental samples (water: 20; swabs: 263) collected from water storage and supply facilities of the Mubarak Al-Kabeer Hospital, Kuwait, were tested by culture and genus-specific PCR for the detection of Legionella. The L. pneumophila isolates were subsequently typed by serology and RAPD-PCR using serotype-specific sera and arbitrary primers, respectively.

RESULTS:

Of the 70 clinical samples, culture yielded 2 (2.9%) whereas genus-specific PCR detected Legionella in 20 (28.6%) samples. The 2 culture-positive specimens were also positive for L.-pneumophila-specific PCR. Testing of swab and water samples by culture and genus-specific PCR yielded 61 (21.6%) and 67 (23.7%) positive samples, respectively. All of the 61 culture-positive samples were also positive by genus-specific PCR and 45 of them were positive for L.-pneumophila-specific PCR. Serological typing of 43 L. pneumophila isolates showed that 8 of these belonged to serotype 1 and 35 to serotype 3; however, RAPD-PCR analyses demonstrated polymorphisms among the isolates of both serotypes.

CONCLUSION:

A higher association between PCR and culture was observed for the environmental samples than for the clinical samples. The application of genus- and species-specific PCRs and RAPD is useful in the detection and typing of Legionella in clinical and environmental samples.

PMID:
18059101
DOI:
10.1159/000109590
[Indexed for MEDLINE]
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