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

Eroglu F,Genc A,Koltas Is

doi:10.4172/2155-9597.1000113

Abstract

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.

Highlights

Cutaneous leishmaniasis (CL) is a skin disease caused by the Leishmania species [7,20]
Specimens from 64 suspected CL patients infected in the Cukurova region, in the southern part of Turkey, were examined by microscopic examination, kinetoplast DNA (kDNA) and mini-exon polymerase chain reaction (PCR) (Table 1)
To be sure about the exact type of species, the PCR products were digested with EaeI for restriction fragment length polymorphism (RFLP). 11 mini-exon PCR according to the microscopic examination; the specificity was 85.2% and the sensitivity was 100% (Table 4)

Summary

Introduction

Cutaneous leishmaniasis (CL) is a skin disease caused by the Leishmania species [7,20]. A clinical evaluation is not enough for a conclusive diagnosis of CL, so the laboratory diagnosis is important for CL diagnosis [27,28]. The traditional diagnostic methods of CL consist of the microscopic examination of smears after Giemsa staining, culture and serological techniques [29]. These techniques are adequate for the diagnosis of CL, but these techniques are not adequate for the discrimination of species due to the morphological similarity of different Leishmania species [11,12]

Methods

Results

Conclusion