The Formosan subterranean termite Coptotermes formosanus Shriaki; (Isoptera: Rhinotermitidae), is a major pest that is spreading throughout the southeastern United States. Morphological identification of worker specimens is not possible using available taxonomic keys based on morphological traits. A multiplex PCR protocol was developed that can differentiate the Formosan subterranean termite from other termite species in a single PCR reaction. This multiplex PCR protocol simplifies previous molecular diagnostic techniques.