Implementation of a PCR-RFLP / Hsp70 to identify and typify Leishmania species in an area endemic to Leishmaniasis Atypical Cutaneous in Honduras

Abstract

Introduction. The atypical cutaneous leishmaniasis is a non-ulcerated variant of the cutaneous leishmaniasis produced by Leishmania (L.) infantum. In Honduras, it is present in the departments of La Paz, Francisco Morazán, Choluteca, El Paraíso, and Valle. The clinical spectrum includes from cutaneous lesions smaller than 10 mm and benign, thru visceral affections tan can compromise the patient’s life. The objective of the present study was to implement the molecular biology technique for the detection and typing of the Leishmania spp in individuals with suspected atypical cutaneous leishmaniasis lesions. Methodology. For the implementation, reference strain DNA was used: Leishmania (L.) infantum chagasi,  leishmania (L.) mexicana, Leishmania (V.) panamensis, Leishmania (V.) braziliensis with an expected product of 1200 base pairs (pb). Hae III, BccI, RsaI, and Mlul restriction enzymes were used for the Leishmania gender specie identification. Film smear clinical samples were selected from departments of Francisco Morazán and Valle. The study was submitted to the Ethics Committee of the Master of Infectious and Zoonotic Diseases. Results: 39 samples were analized (27 from Francisco Morazán and 12 from Valle). Leishmania (L.) infantum was identified in the 100% of the samples (29/29). Conclusions/Recommendations: It was demonstrated that the polimorphism of the hsp70 gene, together with the enzymatic digestion (RFLP) provides good results in the parasite amplification from film smear samples, expediting the molecular identification, being a very useful tool for the Leishmania spp specie typication in areas where this parasite is more prevalent. It is recommended to make a constant epidemiological surveillance to determine the Leishmania specie geographical behavior.