Unsuitable distinction between viable and deadStaphylococcus aureusandStaphylococcus epidermidisby ethidium bromide monoazide

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Comparison 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...

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Ethidium 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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Possible errors in the interpretation of ethidium bromide and PicoGreen DNA staining results from ethidium monoazide-treated DNA.

Nocker and Camper reported that DNA from dead bacterial cells (Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium) treated with ethidium monoazide (EMA) is lost during the DNA extraction procedure (2). This was concluded from the missing or less intense DNA band on an agarose gel stained with ethidium bromide in comparison to the intensities of bands in untreated samples. Acco...

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Enumeration 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.

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Molecular Characterization of Viable Legionella spp. in Cooling Tower Water Samples by Combined Use of Ethidium Monoazide and PCR

Viable Legionella spp. in environmental water samples were characterized phylogenetically by a clone library analysis combining the use of ethidium monoazide and quantitative PCR. To examine the diversity of Legionella spp., six cooling tower water samples and three bath water samples were collected and analyzed. A total of 617 clones were analyzed for their 16S rRNA gene sequences and classifi...

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ژورنال

عنوان ژورنال: Letters in Applied Microbiology

سال: 2009

ISSN: 0266-8254,1472-765X

DOI: 10.1111/j.1472-765x.2009.02585.x