Differentiation of old and new world leishmania species at complex and species levels by PCR.

The variable and conserved sequence boxes of kinetoplast DNA (kDNA) of 11 standard strains of 6 complexes of New and Old World Leishmania were amplified using PCR. Four strains from 2 complexes of Old World Leishmania - L. major (MRHO/IR/64/Nadim-1), with 2 bands at 850 and 620 bp, L. major (MHOM/SU/73/5-ASKH), with a band at 620 bp, L. donovani, with a band at 800 bp and L. infantum, with a band at 650 bp - could be differentiated from each other and from the New World strains, with the exception of L. infantum. Seven Leishmania strains from 4 complexes of New World Leishmania - L. mexicana and L. pifanoi, with a band at 730 bp, L. guyanensis, with 2 bands at 730 and 650 bp, L. peruviana, with a band at 710 bp and L. amazonensis, L. garnhami and L. braziliensis, each with a band at 650 bp - were identified. Of these strains, L. guyanensis and L. peruviana could be differentiated from each other and from the Old World strains. These results show that using PCR amplification of kDNA we could differentiate between New and Old World Leishmania at both complex and strain levels. The amplified kDNA PCR products, together with other techniques, could be useful as a diagnostic tool for the identification of Leishmania species.