Researchers and growers often monitor for the presence of economically important sesiid pests using pheromone traps. These traps often attract more than one species of Sesiidae and specimens often degrade making identification difficult or impossible. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mtDNA cytochrome oxidase I (COI), tRNA leucine, and COII gene using two restriction enzymes, Dra I and Hinf I, provides an accurate method to differentiate eight species of adult clearwing moths commonly found in traps baited with commercially available pheromone lures: Melittia satyriniformis, Paranthrene simulans, Pennisetia marginata, Synanthedon pictipes, S. exitiosa, S. scitula, S. rileyana, and Vitacea polistiformis.