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. 2000 Nov;38(11):4215-8.
doi: 10.1128/JCM.38.11.4215-4218.2000.

Detection of Legionella pneumophila using a real-time PCR hybridization assay

A L Ballard  1 , N K Fry, L Chan, S B Surman, J V Lee, T G Harrison, K J Towner
Affiliations

Detection of Legionella pneumophila using a real-time PCR hybridization assay

A L Ballard et al. J Clin Microbiol. 2000 Nov.

Abstract

A real-time PCR hybridization assay for Legionella pneumophila is described; the assay uses LightCycler (Idaho Technology) methodology to specifically detect 2.5 CFU/reaction, equivalent to 1,000 CFU/liter of starting water sample. The assay, including DNA extraction and confirmation of product identity, is completed within 90 min of receipt of a sample.

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Figures

FIG. 1
FIG. 1
Melting curves for L. pneumophila-specific LightCycler-based PCR. Shown are changes in SYBR green (A) and Cy5 (B) fluorescence (−dF/dT) versus temperature. ——, L. pneumophila serogroup 1; -----, negative control.
FIG. 2
FIG. 2
Quantification of L. pneumophila-specific double-stranded PCR product as measured by SYBR green fluorescence. (A) Log fluorescence versus cycle number (a, 1,000 CFU/reaction; b, 100 CFU/reaction; c, 25 CFU/reaction; d, 2.5 CFU/reaction); (B) crossing points (cycle numbers) of log linear correlations with the noise line band, plotted against the logarithmic concentration of the standards.
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