Development of an Improved Pcr-Ict Hybrid Assay for Direct Detection of Legionellae and Legionella Pneumophila from Cooling Tower Water Specimens
Resource
WATER RESEARCH v.40 n.11 pp.2221-2229
Journal
WATER RESEARCH
Journal Volume
v.40
Journal Issue
n.11
Pages
2221-2229
Date Issued
2006
Date
2006
Author(s)
HORNG, YU-TZE
SOO, PO-CHI
SHEN, BIN-JON
SU, HSUN-PI
WEI, JIUN-RUNG
HSIEH, SHANG-CHEN
HSUEH, PO-REN
LAI, HSIN-CHIH
Abstract
A novelly improved polymerase chian reaction and immunochromatography test (PCR-ICT) hybrid assay comprising traditional multiplex-nested PCR and ICT, (a lateral-flow device) was developed for direct detection of Legionella bacteria from environmental cooling tower samples. The partial 16S rDNA (specific for Legionella spp.) and dnaJ ( specific for Legionella pneumophila) genes from Legionella chromosome were first specifically amplified by multiplex- nested PCR, respectively, followed by detection using ICT strip. Reading of results was based on presence or absence of the two test lines on the strips. Presence of test line 1 indicated existence of Legionella spp. specific 16S rDNA and identified Legionella spp. Presence of test line 2 further indicated existence of dnaJ and thus specifically identified L. pneumophila. In contrast, for non-Legionellae bacteria no test line formation was observed. Results of direct detection of Legionella bacteria and L. pneumophila from water tower specimens by this assay showed 100% sensitivity, and 96.6% and 100% specificity, respectively compared with traditional culture, biochemical and serological identification methods. The PCR-ICT hybrid assay does not require sophisticated equipment and was proved to be practically useful in rapid and direct Legionellae detection from environmental water samples. (c) 2006 Elsevier Ltd. All rights reserved.
Subjects
Legionella pneumophila
PCR-ICT
lateral flow