Identification of cutaneous leishmaniasis agents by nested polymerase chain reaction [Nested-PCR] in Shush city, Khuzestan province, Iran

Leishmaniasis is a common parasitic disease worldwide. Leishmania tropica and L. major are two common cause of cutaneous leishmaniasis in Iran. The aim of this study was determination of the cause of cutaneous leishmaniasis in Shush city, Khouzestan Province, Southwest Iran. One hundred samples were collected from patients at the age of 1-80 year with documented cutaneous leishmaniasis referred to the health centre and a private medical diagnostic laboratory at Shush City. DNA was extracted from slid samples by phenol- chloroform- Isoemil alcohol method, and subjected to Nested-PCR as template. k DNA of the parasites were amplified by CSB1XR and CSB2XF in the first round of PCR and 13Z and Li R primers for the second round. After PCR, electrophoresis of products was performed and 750bp band from L. tropica and 560bp band from L. major were detected. A total of 100 cases comprising 47 females and 53 males were studied. The highest infected age group was under 10 years with a rate of 42% and the lowest rate was 4% at the age group of above 40 years. The results of PCR electrophoresis indicated that 90[90%] cases were L. major and 10 [10%] L. tropica. The predominant species in this area was L. major. It is concluded that Nested PCR is a reliable test for diagnosis and identification of Leishmania species and can apply in epidemiological investigations