Detection of viableEscherichia coliO157:H7 by ethidium monoazide real-time PCR
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چکیده
منابع مشابه
Specific detection of viable Legionella cells by combined use of photoactivated ethidium monoazide and PCR/real-time PCR.
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...
متن کاملEnumeration of viable Listeria monocytogenes cells by real-time PCR with propidium monoazide and ethidium monoazide in the presence of dead cells.
Propidium monoazide (PMA) and ethidium monoazide were used for enumeration of viable Listeria monocytogenes cells in the presence of dead cells. PMA had no antimicrobial effect on L. monocytogenes. Viable cell counts were linearly related to real-time PCR threshold cycle values for PMA-treated cells from planktonic and biofilm sources over a 4-log range.
متن کاملSelective detection of viable seed-borne Acidovorax citrulli by real-time PCR with propidium monoazide
In recent years, use of the DNA-intercalating dye propidium monoazide (PMA) in real-time PCR has been reported as a novel method to detect viable bacteria in different types of samples, such as food, environmental, and microbiological samples. In this study, viable cells of Acidovorax citrulli, the causal agent of bacterial seedling blight and fruit blotch, were selectively detected and differe...
متن کاملComparison of ethidium monoazide and propidium monoazide for the selective detection of viable Legionella cells.
Ethidium monoazide (EMA) and propidium monoazide (PMA) have been utilized for selective PCR amplification of DNA from viable bacterial cells. In this study, we compared the abilities of EMA and PMA, together with real-time PCR, to specifically distinguish dead Legionella cells from viable cells. Several experiments showed that PMA or EMA treatment could specifically prevent the PCR amplificatio...
متن کاملEthidium monoazide for DNA-based differentiation of viable and dead bacteria by 5'-nuclease PCR.
PCR techniques have significantly improved the detection and identification of bacterial pathogens. Even so, the lack of differentiation between DNA from viable and dead cells is one of the major challenges for diagnostic DNA-based methods. Certain nucleic acid-binding dyes can selectively enter dead bacteria and subsequently be covalently linked to DNA. Ethidium monoazide (EMA) is a DNA interc...
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ژورنال
عنوان ژورنال: Journal of Applied Microbiology
سال: 2009
ISSN: 1364-5072,1365-2672
DOI: 10.1111/j.1365-2672.2009.04358.x