Development of a polymerase chain reaction-restriction fragment length polymorphism assay for Leishmania major/Leishmania killicki/Leishmania infantum discrimination from clinical samples, application in a Tunisian focus

Topoisomerase II gene of Leishmania genus was used to develop a molecular tool for detection and species differentiation of Leishmania from clinical samples. Identification was achieved by a polymerase chain reaction followed by digestion with 2 restriction endonucleases BstU1 and Taq1. Despite the relatively low sensitivity, it is able to differentiate between 3 complexes responsible for cutaneous leishmaniasis.