SUMMARYDNA–DNA hybridization techniques have been found to be very useful in the classification and identification of Leishmania parasites. We report here the cloning of a mini-circle from Leishmania strain UR6 in a plasmid and subcloning of the mini-circle fragments in M13 mp9. Clone MLURk32, containing a 560 bp fragment of mini-circle, has been found to have unique specificity. Application of this specific probe in identifying different Leishmania isolates reveals that the probe reacted only with strains of post-kala-azar dermal leishmaniasis but not with strains or isolates of visceral leishmaniasis.