Development and Clinical Evaluation of an Internally Controlled, Single-tube Multiplex Real-time PCR Assay for Detection of Legionella Pneumophila and Other Legionella Species

Overview

Journal J Clin Microbiol

Specialty Microbiology

Date 2003 Sep 6

PMID 12958219

Citations 35

Authors

Kate E Templeton

Sitha A Scheltinga

Peter Sillekens

Jantine W Crielaard

Alje P van Dam

Herman Goossens

Eric C J Claas

Affiliations

Soon will be listed here.

Abstract

A multiplex real-time PCR assay for detection of Legionella pneumophila and Legionella spp. and including an internal control was designed. Legionella species, L. pneumophila, and the internal control were detected simultaneously by probes labeled with 6-carboxy-fluorescein, hexachlorofluorescein, and indodicarbocyanine, respectively. Therefore, no postamplification analysis was required in order to distinguish the targets. The sensitivity of both assays was 2.5 CFU/ml, and from analysis of 10 culture-positive and 74 culture-negative samples from patients investigated for legionellosis, 100% agreement was observed by both assays in comparison to culture. Four additional positives were found by the multiplex real-time PCR assay in the Legionella culture-negative samples.

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