Plectropomus leopardus , Promicrops lanceolatus and Cromileptes altivelis are high-value groupers in chinese aquatic product market and are substituted by cheaper ones sometimes due to economic profit. In this study, real-time PCR methods for identification of these groupers were developed. Species-specific primers and probes were designed on the basis of cytochrome oxidase I (COI) mitochondrial DNA sequences of groupers and other fishes. The specificity of the real time PCR assays were tested with a total of 51 specimens of fish, meat and poultry. Amplifications were observed in specific species only. The methods developed were accurate and rapid and offered the potential to detect fraudulent or unintentional mislabeling of these species in routine seafood authentication analysis. [ABSTRACT FROM AUTHOR]