Table 1.
East African L. donovani strains for which sequences were determined, with GenBank accession numbers.
Table 2.
GenBank sequences used in comparisons.
Figure 1.
Leishmania rK39 and HASPB antigen repeats and the PCR primer target sequences.
Repetitive coding regions depicted as filled boxes, PCR primers underlined, and 5′ and 3′ binding positions with amplicons are indicated by dashed lines. [A] Kinesin gene comparison for primer design (* = non-conserved nucleotide). [B] HASPB1 GenBank sequence displays 22× perfect 14aa repeats. [C] HASPB2 GenBank sequence displays 3 imperfect repeats.
Figure 2.
Multiple amplicons corresponding to kinesin tandem repeats are produced by PCR primers LdonK39F and LdonK39R.
Amplifications from strains HU3 (LV9), Hussen, and UGX-MARROW, are depicted. Major amplicon sizes differ by 117 bp, the size of the nucleotide sequence encoding the 39aa repeat in the kinesin gene; mk = Hyperladder I (Bioline).
Table 3.
rK39 polymorphism among East African L. donovani strains and in comparison with the LcKin diagnostic rK39 sequence.
Figure 3.
Polymorphisms among seven rK39 repeats of East African and South Asian strains show region-specific substitutions.
Changes between charged and non-charged residues are underlined. No residue entered at a particular site indicates conservation of that residue with the corresponding residue of diagnostic rK39. Where two alternative residues are indicated in smaller text these are not necessarily always within the same strain, for clarification compare with Table 3. Region-specific polymorphisms in each repeat are boxed (East Africa) or circled (South Asia).
Figure 4.
Predicted (1064 bp, 260 bp) amplicons and unpredicted (∼400–500 bp) amplicons with HASPB PCR primers LdonHASPBfor and LdonHASPBrev.
Amplifications from strains HU3 (LV9), Hussen, UGX-MARROW, and LRC-L57, are depicted; mk = Hyperladder I.
Table 4.
Repeat sequences of unexpected amplicons.