Objectives: Accurate identification and quantification of oral pathogenic bacteria could be an invaluable tool to aid the treatment of oral diseases such as caries and periodontal diseases. Conventional methods, such as counting colony forming unit (CFU) and colorimetric methods, suffer various limitations besides being time-consuming and cumbersome. Therefore, aim of the present study was to develop a multiplex Taqman assay for simultaneous detection of a wide range of oral pathogens and to validate the assay using clinical samples. Methods: The following bacterial species were used in the study: Streptococcus mutants, Streptococcus sobrinus, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Fusobacteirum necleatum and Enterococcus faecalis. Bacterial DNA corresponding to 1 x 103 -1 x 108 cells was prepared from bacterial cultures to generate a standard curve. Species-specific Taqman probes and primers were designed using bioinformatics tools. Thereafter, multiplex PCR array was optimized to detect three bacterial species in a single well and further validated using clinical samples. Results: Taqman primer-probe pairs designed for each species showed excellent specificity without cross-reacting with other species. Multiplex Taqman PCR formula was able to detect three species such as P. gingivalis, A. actinomycetemcomitans and S. mutans in a single well without competitive inhibition. Clinical samples showed that Taqman probes were more specific and sensitive to detect aforementioned pathogen than conventional techniques. Conclusions: The present quantitative multiplex Taqman PCR array developed for identification and quantification of oral pathogens may serve as a useful diagnostic and prognostic tool for oral infectious diseases (Supported by HKU 201007160018 & 201007159003 to CJS).