Table 1.
Leishmania strains isolated in Ecuador.
Table 2.
Primer sequences used in this study.
Fig 1.
PCR-RFLP analyses of mpi gene fragments from 6 Leishmania species in Ecuador.
PCR amplification was performed with leishmanial mpi gene-specific primers, and PCR products were digested with (A) HaeIII and (B) HpaI, and resulting restriction fragment patterns were analyzed by agarose gel electrophoresis. 1. L. (V.) guyanensis, 2. L. (V.) panamensis, 3. L. (V.) braziliensis, 4. L. (V.) naiffi, 5. L. (L.) major-like, 6. L. (L.) mexicana.
Table 3.
Fragment size of leishmanial mpi and 6pgd genes generated by digestion with selected restriction enzymes.
Fig 2.
PCR-RFLP analyses of 6pgd gene fragments from 6 Leishmania species in Ecuador.
PCR amplification was performed with leishmanial 6pgd gene-specific primers, and PCR products were digested with (A) Bsp1286I and (B) HinfI, and resulting restriction fragment patterns were analyzed by agarose gel electrophoresis. 1. L. (V.) guyanensis, 2. L. (V.) panamensis, 3. L. (V.) braziliensis, 4. L. (V.) naiffi, 5. L. (L.) major-like, 6. L. (L.) mexicana.
Fig 3.
Differentiation between L. (V.) guyanensis and L. (V.) panamensis by PCR-RFLP of the hsp70 gene fragment.
PCR amplification was performed with hsp70 gene-specific primers and the PCR products were digested with BccI. 1. L. (V.) guyanensis, 2. L. (V.) panamensis, 3. a hybrid of L. (V.) guyanensis and L. (V.) panamensis.
Fig 4.
Geographic distribution of Leishmania species in Ecuador identified by PCR-RFLP analyses targeting multiple nuclear genes.
The dark gray areas show the Andean plateau (>1,000 m altitude), and the light gray areas show highland jungle or Andean slopes (400–1,000 m elevation). (Adapted from a map available at http://english.freemap.jp/).
Fig 5.
Differentiation between L. (V.) guyanensis and L. (V.) braziliensis by PCR-RFLP of mpi and 6pgd gene fragments.
A, B. PCR amplification was performed with mpi gene- or 6pgd gene-specific primers and the PCR products were digested with HaeIII (A) or Bsp1286I (B), respectively. 1. L. (V.) guyanensis, 2. L. (V.) braziliensis, 3. a hybrid of L. (V.) guyanensis and L. (V.) braziliensis.
Table 4.
Comparison of Leishmania species identification in Ecuador between cyt b sequence analysis and PCR-RFLP analyses of nuclear DNAs.