| Abstract: | Leishmania donovani is the known causative agent of both cutaneous(CL) and visceral leishmaniasis in Sri Lanka. CL is considered to be under-reportedpartly 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 samplesfrom patients presented with suspected lesions in 2010. Both,Leishmaniagenus-specific JW11/JW12 KDNA and LITSR/L5.8S internaltranscribed spacer (ITS)1 PCR assays detected 92% (35/38) of the samples whereas aKDNA assay specific forL. donovani (LdF/LdR) detected only 71%(27/38) of samples. All positive samples showed a L. donovanibanding pattern upon HaeIII ITS1 PCR-restriction fragment length polymorphismanalysis. PCR assay specificity was evaluated in samples containingMycobacterium tuberculosis, Mycobacteriumleprae, and human DNA, and there was no cross-amplification in JW11/JW12and 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 inM. tuberculosis samples. In conclusion, it was successfully shownin this study that it is possible to diagnose Sri Lankan CL with high accuracy, togenus and species identification, using Leishmania DNA PCRassays. |
