RRC ID 5361
著者 Chang B, Sugiyama K, Taguri T, Amemura-Maekawa J, Kura F, Watanabe H.
タイトル Specific detection of viable Legionella cells by combined use of photoactivated ethidium monoazide and PCR/real-time PCR.
ジャーナル Appl Environ Microbiol
Abstract Legionella organisms are prevalent in manmade water systems and cause legionellosis in humans. A rapid detection method for viable Legionella cells combining ethidium monoazide (EMA) and PCR/real-time PCR was assessed. EMA could specifically intercalate and cleave the genomic DNA of heat- and chlorine-treated dead Legionella cells. The EMA-PCR assay clearly showed an amplified fragment specific for Legionella DNA from viable cells, but it could not do so for DNA from dead cells. The number of EMA-treated dead Legionella cells estimated by real-time PCR exhibited a 10(4)- to 10(5)-fold decrease compared to the number of dead Legionella cells without EMA treatment. Conversely, no significant difference in the numbers of EMA-treated and untreated viable Legionella cells was detected by the real-time PCR assay. The combined assay was also confirmed to be useful for specific detection of culturable Legionella cells from water samples obtained from spas. Therefore, the combined use of EMA and PCR/real-time PCR detects viable Legionella cells rapidly and specifically and may be useful in environmental surveillance for Legionella.
巻・号 75(1)
ページ 147-53
公開日 2009-1-1
DOI 10.1128/AEM.00604-08
PII AEM.00604-08
PMID 18978073
PMC PMC2612233
MeSH Azides / metabolism* Humans Legionella / genetics Legionella / isolation & purification* Male Microbial Viability Polymerase Chain Reaction / methods* Sensitivity and Specificity Staining and Labeling / methods* Water Microbiology*
IF 4.016
引用数 55
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY
リソース情報
一般微生物 JCM7516 JCM10698 JCM20691 JCM1379 JCM2506 JCM13232 JCM13912 JCM20371