The　main　disadvantage　of　DNA-based　quantification　methods　is　their　inability　to　distinguish　between　viable　and　non-viable　cells,　as　dead　cells　may　also　retain　a　significant　amount　of　DNA.　To　overcome　this　limitation,　propidium　monoazide　(PMA)　treatment　was　applied　to　effectively　prevent　amplification　of　DNA　from　dead　cells.　The　objectives　of　this　study　were　to　develop　and　evaluate　an　accurate　detection　method　that　combines　PMA　staining　with　real-time　qPCR　to　quantify　viable　dominant　microorganisms　during　the　traditional　brewing　of　Hong　Qu　glutinous　rice　wine.　Firstly,　three　methodological　factors　(concentration　of　PMA,　incubation　time　and　photoactivation　time)　were　optimized　for　representative　strains　of　different　microbial　species.　For　the　tested　microbial　strains　(Monascus　purpureus,　Saccharomyces　cerevisiae　and　Lactobacillus　plantarum),　a　treatment　of　25　mu　M　PMA　for　20　min　incubation　followed　by　10　min　photoactivation　was　considered　optimal　to　achieve　a　compromise　between　minimal　impact　on　intact　cells　and　maximal　signal　reduction　for　the　compromised　cells.　Compared　with　qPCR　without　PMA　treatment,　results　of　PMA-qPCR　showed　that　the　optimized　PMA　pretreatment　can　effectively　exclude　the　dead　microbial　cells　without　any　negative　effects　on　the　intact　cells.　The　concentrations　of　viable　cells　calculated　by　PMA-qPCR　were　comparable　(the　same　order　of　magnitude　was　obtained)　with　those　obtained　by　culture-based　method.　Finally,　the　developed　PMA-qPCR　methods　were　successfully　applied　to　monitor　the　total　bacteria,　LAB,　fungi,　yeasts,　M.　purpureus,　S.　cerevisiae　and　L.　plantarum　during　the　traditional　brewing　of　Hong　Qu　glutinous　rice　wine.　Both　of　the　total　bacterial　and　fungal　populations　increased　significantly　at　the　early　brewing　stage,　but　they　decreased　at　the　later　brewing　stage.　PMA-qPCR　results　revealed　that　the　predominant　microorganisms　varied　in　different　brewing　phases.　L.　plantarum　and　S.　cerevisiae　grew　significantly　and　became　the　predominant　bacterial　and　fungal　species　at　the　final　stage　of　traditional　brewing　process,　respectively.　This　study　standardized　the　PMA-qPCR　parameters　for　dominant　microorganisms　associated　with　the　traditional　brewing　of　Hong　Qu　glutinous　rice　wine,　which　would　show　great　value　in　scientifically　understanding　of　the　brewing　mechanism　and　provide　useful　information　for　the　selection　of　beneficial　microbial　strains　to　improve　wine　quality.　(C)　2016　Published　by　Elsevier　Ltd.