{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T16:13:09Z","timestamp":1786464789296,"version":"build-2736575974"},"reference-count":55,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T00:00:00Z","timestamp":1786406400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p>\n                    Accurate quantification of viable bacteria is essential for ensuring product safety and quality in the food, pharmaceutical, and cosmetics industries. However, conventional plate culture methods are time-consuming and fail to detect viable but non-culturable (VBNC) cells, which frequently occur under environmental stress or preservation conditions and can influence product stability and safety. Accordingly, the need for a quantification method that accounts for VBNC cells has increased. In this study, we optimized and validated a viability-selective quantification method by combining propidium monoazide (PMA), a membrane-impermeable dye, with quantitative real-time polymerase chain reaction (qPCR).\n                    <jats:italic>Staphylococcus aureus<\/jats:italic>\n                    BF00309,\n                    <jats:italic>Staphylococcus epidermidis<\/jats:italic>\n                    CICARIA, and\n                    <jats:italic>Lactobacillus jensenii<\/jats:italic>\n                    KCTC 5194 were analyzed to validate the method. PMA treatment effectively suppressed DNA amplification in dead cells, resulting in significantly higher\n                    <jats:italic>C<\/jats:italic>\n                    <jats:sub>q<\/jats:sub>\n                    values than in untreated controls. Strong linear correlations between\n                    <jats:italic>C<\/jats:italic>\n                    <jats:sub>q<\/jats:sub>\n                    values and colony-forming units (CFU) obtained by plate counting were observed for all strains (\n                    <jats:italic>R<\/jats:italic>\n                    <jats:sup>2<\/jats:sup>\n                    &amp;gt; 0.99), enabling the establishment of a\n                    <jats:italic>C<\/jats:italic>\n                    <jats:sub>q<\/jats:sub>\n                    \u2013CFU regression model for estimating viable cell numbers. The derived model successfully quantified membrane-intact viable cells, which may include potential VBNC-like populations, and showed high agreement with culture-based methods. Moreover, selective detection of target bacteria was achieved in mixed samples containing high concentrations of background microorganisms, with a stable detection limit of approximately 10\n                    <jats:sup>3<\/jats:sup>\n                    CFU\/mL. These results demonstrate that the optimized PMA-qPCR method provides a rapid complementary tool to conventional culture-based quantification and serves as a promising preliminary screening approach with strong potential for future applications in skin microbiome research and the evaluation of live bacteria-based products, pending further validation in complex product matrices.\n                  <\/jats:p>","DOI":"10.3389\/fmicb.2026.1873833","type":"journal-article","created":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T13:57:16Z","timestamp":1786456636000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Application-focused validation of a viability-selective PMA-qPCR assay for selected skin-associated and cosmetic-relevant bacteria"],"prefix":"10.3389","volume":"17","author":[{"given":"Sejin","family":"Cheon","sequence":"first","affiliation":[{"name":"COSMAX BTI R&I Center, Bundang-Gu","place":["Seongnam, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jimin","family":"Yoo","sequence":"additional","affiliation":[{"name":"Department of Microbiology, College of Bio-convergence, Dankook University","place":["Cheonan, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunseok","family":"Oh","sequence":"additional","affiliation":[{"name":"Department of Microbiology, College of Bio-convergence, Dankook University","place":["Cheonan, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haeun","family":"Lee","sequence":"additional","affiliation":[{"name":"COSMAX BTI R&I Center, Bundang-Gu","place":["Seongnam, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dong-geol","family":"Lee","sequence":"additional","affiliation":[{"name":"COSMAX BTI R&I Center, Bundang-Gu","place":["Seongnam, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seunghyun","family":"Kang","sequence":"additional","affiliation":[{"name":"COSMAX BTI R&I Center, Bundang-Gu","place":["Seongnam, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunho","family":"Park","sequence":"additional","affiliation":[{"name":"COSMAX BTI R&I Center, Bundang-Gu","place":["Seongnam, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seyoung","family":"Mun","sequence":"additional","affiliation":[{"name":"Department of Microbiology, College of Bio-convergence, Dankook University","place":["Cheonan, Republic of Korea"]},{"name":"Center for Bio-Medical Engineering Core Facility, Dankook University","place":["Cheonan, Republic of Korea"]},{"name":"Department of Cosmedical Materials, College of Bio-convergence, Dankook University","place":["Cheonan, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Young Mok","family":"Heo","sequence":"additional","affiliation":[{"name":"COSMAX BTI R&I Center, Bundang-Gu","place":["Seongnam, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyudong","family":"Han","sequence":"additional","affiliation":[{"name":"Department of Microbiology, College of Bio-convergence, Dankook University","place":["Cheonan, Republic of Korea"]},{"name":"Center for Bio-Medical Engineering Core Facility, Dankook University","place":["Cheonan, Republic of Korea"]},{"name":"Department of Cosmedical Materials, College of Bio-convergence, Dankook University","place":["Cheonan, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,8,11]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/s00284-023-03526-3","article-title":"Quality management of probiotics: Ensuring safety and maximizing health benefits.","volume":"81","author":"Ahire","year":"2024","journal-title":"Curr. 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