Literature DB >> 18978073

Specific detection of viable Legionella cells by combined use of photoactivated ethidium monoazide and PCR/real-time PCR.

Bin Chang1, Kanji Sugiyama, Toshitsugu Taguri, Junko Amemura-Maekawa, Fumiaki Kura, Haruo Watanabe.   

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.

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Year:  2008        PMID: 18978073      PMCID: PMC2612233          DOI: 10.1128/AEM.00604-08

Source DB:  PubMed          Journal:  Appl Environ Microbiol        ISSN: 0099-2240            Impact factor:   4.792


  25 in total

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  14 in total

1.  Specific and rapid enumeration of viable but nonculturable and viable-culturable gram-negative bacteria by using flow cytometry.

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8.  Mycobacterium avium complex in day care hot water systems, and persistence of live cells and DNA in hot water pipes.

Authors:  Annette S Bukh; Peter Roslev
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