Analysis of Kinetoplast DNA from Mexican Isolates ofLeishmania (L.) mexicana

Abstract

This study analyzed DNA minicircles of Mexican isolates ofL. (Leishmania) mexicanato look for genetic differences between strains isolated from patients with diffuse cutaneous (DCL) and localized (LCL) leishmaniasis. The kDNA was analyzed using polymerase chain reaction (PCR), restriction fragment polymorphism analysis of the PCR products (PCR-RFLP) and the PCR products were sequenced. In the PCR with primers specific for the subgenusLeishmania, the Mexican isolates gave higher amplification products than the otherL. mexicanacomplex strains and with specific primers for theL. mexicanacomplex they were poorly amplified. In the PCR-RFLP analysis with theEco RV,Hae III, andMbo Iendonucleases, the Mexican isolates displayed similar restriction patterns, but different from the patterns of the other members of theL. mexicanacomplex. In the phylogenetic tree constructed, the kDNA sequences of the Mexican clones formed two groups including sequences of LCD or LCL clones, apart from the otherL. mexicanacomplex members. These results suggest that the kDNA minicircles of the Mexican isolates are more polymorphic than the kDNA of other members of theL. mexicanacomplex and have different recognition sites for the restriction enzymes used in this study.