A molecular procedure incorporating polymerase chain reaction (PCR) of the COI gene and restriction endonuclease digestion of PCR products was used to distinguish Peristenus howardi (Hymenoptera: Braconidae) from four other Peristenus species. Non-solvent extraction of parasite DNA using a commercially available kit proved to be very effective in producing amplifiable template. Use of SfcI endonuclease produced restriction fragments with banding patterns in agarose gel electrophoresis that readily separated P. howardi, P. digoneutis, P. conradi, P. pallipes, and P. pseudopallipes. However, while the restriction fragment banding patterns of both P. pallipes and P. pseudopallipes were easily distinguishable from the other Peristenus species, they could not be reliably separated from one another. This molecular procedure can be used in applied and ecological research to better understand the role of P. howardi in the Peristenus-Lygus parasite-host system within the Pacific Northwest. Consensus sequences of our amplimers for all five Peristenus spp. are deposited in GenBank under accession numbers AY626370, AY626371, AY626372, AY626373, and AY626374.