{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,2]],"date-time":"2025-11-02T16:35:44Z","timestamp":1762101344140},"reference-count":36,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2012,9,7]],"date-time":"2012-09-07T00:00:00Z","timestamp":1346976000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J of Periodontal Research"],"published-print":{"date-parts":[[2013,4]]},"abstract":"<jats:sec><jats:title>Background and Objectives<\/jats:title><jats:p>One of the major disadvantages of <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content>\u2010based microbial diagnostics is their inability to differentiate <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> between viable and dead microorganisms, which could be important when studying etiologically relevant pathogens. The aim of this investigation was to optimize a method for the selective detection and quantification of only viable <jats:italic><jats:styled-content style=\"fixed-case\">A<\/jats:styled-content>ggregatibacter\u00a0actinomycetemcomitans<\/jats:italic> and <jats:italic><jats:styled-content style=\"fixed-case\">P<\/jats:styled-content>orphyromonas\u00a0gingivalis<\/jats:italic> cells by combining quantitative real\u2010time polymerase chain reaction (q<jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content>) and propidium monoazide (<jats:styled-content style=\"fixed-case\">PMA<\/jats:styled-content>).<\/jats:p><\/jats:sec><jats:sec><jats:title>Material and Methods<\/jats:title><jats:p>Three different concentrations of <jats:styled-content style=\"fixed-case\">PMA<\/jats:styled-content> (10, 50 or 100\u00a0\u03bc<jats:sc>m<\/jats:sc>) were added to suspensions of 10<jats:sup>6<\/jats:sup>\u00a0(CFU)\/mL of viable\/dead <jats:italic><jats:styled-content style=\"fixed-case\">A<\/jats:styled-content>.\u00a0actinomycetemcomitans<\/jats:italic> and <jats:italic><jats:styled-content style=\"fixed-case\">P<\/jats:styled-content>.\u00a0gingivalis<\/jats:italic> cells. After <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> isolation, q<jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> was carried out using specific primers and probes for the tested bacteria. <jats:styled-content style=\"fixed-case\">PMA<\/jats:styled-content> was further tested with different mixtures containing varying ratios of viable and dead cells. The efficacy of <jats:styled-content style=\"fixed-case\">PMA<\/jats:styled-content> to detect viable\/dead cells was tested by analysis of variance.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>For these specific bacterial pathogens, 100\u00a0\u03bc<jats:sc>m <\/jats:sc><jats:styled-content style=\"fixed-case\">PMA<\/jats:styled-content> resulted in a significant reduction of q<jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> amplification with dead cells (10<jats:sup>6<\/jats:sup>\u00a0CFU\/mL), while with viable cells no significant inhibition was detected. <jats:styled-content style=\"fixed-case\">PMA<\/jats:styled-content> was also effective in detecting selectively viable cells by q<jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> detection, when mixtures of varying ratios of viable and dead bacteria were used.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>This study demonstrated the efficiency of <jats:styled-content style=\"fixed-case\">PMA<\/jats:styled-content> for differentiating viable and dead <jats:italic><jats:styled-content style=\"fixed-case\">A<\/jats:styled-content>.\u00a0actinomycetemcomitans<\/jats:italic> and <jats:italic><jats:styled-content style=\"fixed-case\">P<\/jats:styled-content>.\u00a0gingivalis<\/jats:italic> cells. This method of <jats:styled-content style=\"fixed-case\">PMA<\/jats:styled-content>\u2010q<jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> may be useful for monitoring new antimicrobial strategies and for assessing the pathogenic potential of <jats:italic><jats:styled-content style=\"fixed-case\">A<\/jats:styled-content>.\u00a0actinomycetemcomitans<\/jats:italic> and <jats:italic><jats:styled-content style=\"fixed-case\">P<\/jats:styled-content>.\u00a0gingivalis<\/jats:italic> in different oral conditions when using molecular diagnostic methods.<\/jats:p><\/jats:sec>","DOI":"10.1111\/j.1600-0765.2012.01522.x","type":"journal-article","created":{"date-parts":[[2012,9,7]],"date-time":"2012-09-07T13:23:00Z","timestamp":1347024180000},"page":"213-220","source":"Crossref","is-referenced-by-count":23,"title":["Analysis of viable vs. dead <i><scp>A<\/scp>ggregatibacter actinomycetemcomitans<\/i> and <i><scp>P<\/scp>orphyromonas gingivalis<\/i> using selective quantitative real\u2010time <scp>PCR<\/scp> with propidium monoazide"],"prefix":"10.1111","volume":"48","author":[{"given":"M. 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