Identification of Causative Species in Cutaneous Leishmaniasis Patients Using PCR-RFLP

Cutaneous leishmaniasis (CL) in Cukurova located in the southern part of Turkey is a public health problem. We assessed the efficiency of a PCR method in establishing the diagnosis of CL. We used two different targets, kinetoplast DNA (kDNA) for diagnosis and a mini-exon gene for species typing. 64 smear samples were taken from clinically CL suspected cases. The DNA was amplified kinetoplast DNA (kDNA) by using the polymerase chain reaction (PCR) with universal primers 13A-13B, specific for the Leishmania genus and DNA was amplified in the mini-exon region by PCR with Fme-Rme primers, specific for Leishmania species. We compared the sensitivity and specificity of conventional microscopy with kDNA and mini-exon PCR. kDNA PCR was found to have a specificty of 58.8% and a sensitivity of 100%, respectively. Furthermore, we performed a restriction fragment length polymorphism (RFLP) analysis on the PCR products of mini-exon for the genotyping of Leishmania species. Interestingly, the PCR-RFLP result showed that 31.5% of the isolates had Leishmania infantum (L.infantum) in CL cases without visceral leishmaniasis (VL) history.