hsp70-nested-PCR-RFLP for Leishmania species identification in clinical samples of ACL. SYBR-safe-stained 2% agarose gels showing HaeIII (lanes 2 to 12) and BccI (lanes 13 to 23) enzymatic digestions of hsp70-nested-PCR products, using as a template the genomic DNA of the following reference strains: lane 2 and 13, L. (V.) guyanensis (MHOM/GF/79/LEM85); lanes 3 and 14, L. (V.) panamensis (MHOM/PA/71/LS94); lanes 4 and 15, L. (V.) braziliensis (MHOM/BR/75/M2903); lanes 5 and 16, L. (L.) amazonensis (MHOM/BR/73/M2269); lanes 6 and 17, L. (L.) mexicana (MHOM/BZ/82/BEL 21); or, using as a template the genomic DNA from different types of clinical samples: lanes 7 and 18, biopsy specimen from patient 21 identified as L. (V.) braziliensis; lanes 8 and 19, Giemsa-stained smear from patient 23 identified as L. (V.) panamensis; lanes 9 and 20, Giemsa-stained smear from patient 15 identified as L. (V.) guyanensis; lanes 10 and 21, biopsy specimen from patient 60 identified as L. (V.) braziliensis; lanes 11 and 22, Giemsa-stained smear from patient 24 identified as L. (V.) panamensis; lanes 12 and 23, biopsy specimen from patient 63 identified as L. (V.) guyanensis; lane 1, size ruler, 50 bp; lane 24, size ruler, 100 bp.