Fig. 5

LOD determination based on PCR amplification of serially diluted DNA from culture promastigotes of Leishmania parasites. DNA was extracted from culture promastigotes of different reference strains of L. major, L. tropica and L. infantum. Serially 10-fold diluted amounts of 100, 10, 1, 0.1, and 0.01 pg of genomic DNAs were spiked into the reactions and the PCR amplification was performed as described with universal primers UNIL-IR-P and UNIL-IR-M. Human skin DNs were also added to each reaction for evaluation of possible interference. The different possible combinations of 3 species were also considered and included in the PCR schedule. PCR products were subjected to electrophoresis on a 1.5% agarose gel. Lanes 1–5: L. major DNA with 100 to 0.01 pg/µl concentrations, respectively; Lanes 6–9: L. tropica DNA with 100 to 0.1 pg/µl concentrations, respectively; Lanes 10–13 L. infantum DNA with 100 to 0.1 pg/µl concentrations, respectively; Lane 14: non-template negative control. Lane L: 100 bp DNA ladder