Abstract: | Leishmania donovani is the known causative agent of both cutaneous
(CL) and visceral leishmaniasis in Sri Lanka. CL is considered to be under-reported
partly due to relatively poor sensitivity and specificity of microscopic diagnosis.
We compared robustness of three previously described polymerase chain reaction (PCR)
based methods to detectLeishmania DNA in 38 punch biopsy samples
from patients presented with suspected lesions in 2010. Both,
Leishmaniagenus-specific JW11/JW12 KDNA and LITSR/L5.8S internal
transcribed spacer (ITS)1 PCR assays detected 92% (35/38) of the samples whereas a
KDNA assay specific forL. donovani (LdF/LdR) detected only 71%
(27/38) of samples. All positive samples showed a L. donovani
banding pattern upon HaeIII ITS1 PCR-restriction fragment length polymorphism
analysis. PCR assay specificity was evaluated in samples containing
Mycobacterium tuberculosis, Mycobacterium
leprae, and human DNA, and there was no cross-amplification in JW11/JW12
and LITSR/L5.8S PCR assays. The LdF/LdR PCR assay did not amplify M.
leprae or human DNA although 500 bp and 700 bp bands were observed in
M. tuberculosis samples. In conclusion, it was successfully shown
in this study that it is possible to diagnose Sri Lankan CL with high accuracy, to
genus and species identification, using Leishmania DNA PCR
assays. |