| RRC ID |
5361
|
| Author |
Chang B, Sugiyama K, Taguri T, Amemura-Maekawa J, Kura F, Watanabe H.
|
| Title |
Specific detection of viable Legionella cells by combined use of photoactivated ethidium monoazide and PCR/real-time PCR.
|
| Journal |
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.
|
| Volume |
75(1)
|
| Pages |
147-53
|
| Published |
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
|
| Times Cited |
55
|
|
WOS Category
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
MICROBIOLOGY
|
| Resource |
| General Microbes |
JCM7516
JCM10698
JCM20691
JCM1379
JCM2506
JCM13232
JCM13912
JCM20371 |
