A simple and reliable technique was developed to distinguish Phlebotomine s
andflies by restriction fragment length polymorphism of PCR-amplified (PCR-
RFLP) 18S rDNAs. Seven morphologically identified sandflies species from se
veral localities of Greece and Cyprus were studied, and specific patterns w
ere developed by double digesting amplified 18S rDNAs with HpaII and RsaI.
Three additional species of the subgenus Larroussius were distinguished by
a second double digestion with AccI and BanI. We have successfully applied
the method on samples in which morphological characters were badly distingu
ished due to poor storage conditions and in larval stages.