Genotyping the causative agents of cutaneous leishmaniasis in Ahvaz southwest of Iran

With sand flies as the main vectors, Leishmania species cause ancient zoonotic diseases called leishmaniasis. Iran is an endemic country regarding cutaneous leishmaniasis. A number of 100 smear slides were collected from cutaneous lesions referred to Ahvaz health centers. The DNA was extracted and ITS1–PCR using LITSR and L5.8S primer pair was performed to detect the genus Leishmania. Then, enzymatic digestion of PCR products was done by HaeIII (species detection), TaqI (strain detection), DpnI and HpaII (mutation assessment). Furthermore, 50 samples were sent for sequencing. Microscopic examination showed amastigote form in all 100 slides. Also, molecular identification confirmed the infection of all cases to Leishmania genus, representing a 350 bp band. HaeIII digestion yielded 150 and 200 bp bands, indicating Leishmania major, while 130 and 200 bp fragments following TaqI digestion suggested A1 strain of the parasite. Moreover, no likely mutations was detected in ITS1 fragment of obtained parasites using DpnI (140 and 200 bp digestion) and HpaII (without digestion). The sequencing result also was consistent with our findings, having 100% homology to A1 strain sequence (AY550178). Leishmania major A1 strain was the predominant species in clinical samples of Ahvaz. Nevertheless, future researches should address the parasite strains in other foci and hosts of epidemiological significance.