Leishmania donovani complex (Kinetoplastida, Trypanosomatidae): comparison of deoxyribonucleic acid based techniques for typing of isolates from Ethiopia.
In Ethiopia, visceral leishmaniasis (VL) is an increasing public health concern. Recently, a new outbreak of VL claimed the lives of hundreds of Ethiopians. Mapping its distribution and the identification of the causative Leishmania species is important for proper use of resources and for control planning. The choice of appropriate typing technique is the key for determining the infecting species. Here we compared three deoxyribonucleic acid (DNA) based markers. We used, for the first time, cpbE and cpbF (cpbE/F) PCR-RFLP and demonstrated that it clearly differentiates Leishmania donovani from Leishmania infantum. The cpbE/F PCR-RFLP gave identical banding pattern for all L. donovani strains irrespective of their geographic origin. With the K26 (primers) PCR-RFLP, the L. donovani strains gave a banding pattern different from L. infantum and showed variation with geographic origin. The Ethiopian isolates typed as L. donovani by the PCR-RFLP of the cpbE/F (gene) and K26 (primers) showed two types of patterns with the T2/B4 (primers) PCR-RFLP; one group with L. infantum-like and the other L. donovani-like pattern. Phylogenetic analysis using cpbE/F sequences showed variation with geographic origin of strains and the African strains of L. donovani are more distantly related to L. infantum. Moreover, the Ethiopian isolates were seen to be closely related to the Sudanese, Kenyan and Indian strains. Thus, we recommend the use of more than one marker to study the population genetics of L. donovani complex.
Gadisa
E
E
Armauer Hansen Research Institute, P.O. Box 1005, Jimma Road, ALERT Campus, Addis Ababa, Ethiopia. endalamawgadisa@yahoo.com
Kuru
T
T
Genet
A
A
Engers
H
H
Aseffa
A
A
Gedamu
L
L
D016054:DNA, Protozoan / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D005002:Ethiopia / Q000453:epidemiology; D005819:Genetic Markers; D005828:Genetics, Population; D006801:Humans; D007893:Leishmania donovani / Q000145:classification* / Q000235:genetics; D007898:Leishmaniasis, Visceral / Q000453:epidemiology / Q000469:parasitology*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D016415:Sequence Alignment
D016054:DNA, Protozoan; D005819:Genetic Markers