title: Polymerase chain reaction detection of Leishmania DNA in skin biopsy samples in Sri Lanka where the causative agent of cutaneous leishmaniasis is Leishmania donovani.
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 detect Leishmania DNA in 38 punch biopsy samples from patients presented with suspected lesions in 2010. Both, Leishmania genus-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.
authors.affiliation: Department of Parasitology, Faculty of Medical Sciences, University of Sri Jayewardenepura, Nugegoda, Sri Lanka.
authors.forename: Shalindra
authors.initials: S
authors.lastname: Ranasinghe
authors.affiliation: Department of Parasitology, Faculty of Medical Sciences, University of Sri Jayewardenepura, Nugegoda, Sri Lanka.
authors.forename: Renu
authors.initials: R
authors.lastname: Wickremasinghe
authors.affiliation: Dermatology Unit, Teaching Hospital Anuradhapura, Anuradhapura, Sri Lanka.
authors.forename: Sanjeeva
authors.initials: S
authors.lastname: Hulangamuwa
authors.affiliation: Dermatology Unit, National Hospital of Sri Lanka, Colombo, Sri Lanka.
authors.forename: Ganga
authors.initials: G
authors.lastname: Sirimanna
authors.affiliation: Dermatology Unit, Teaching Hospital Anuradhapura, Anuradhapura, Sri Lanka.
authors.forename: Nandimithra
authors.initials: N
authors.lastname: Opathella
authors.affiliation: Centre for Applied Entomology and Parasitology, Keele University, Staffordshire, UK.
authors.forename: Rhaiza D C
authors.initials: RD
authors.lastname: Maingon
authors.affiliation: Department of Chemistry, Faculty of Science, University of Colombo, Colombo, Sri Lanka.
authors.forename: Vishvanath
authors.initials: V
authors.lastname: Chandrasekharan
mesh_terms: D001706:Biopsy; D017931:DNA Primers; D016054:DNA, Protozoan / Q000302:isolation & purification*; D006801:Humans; D007893:Leishmania donovani / Q000235:genetics*; D016773:Leishmaniasis, Cutaneous / Q000469:parasitology* / Q000473:pathology; D058069:Neglected Diseases / Q000469:parasitology; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D012867:Skin / Q000469:parasitology* / Q000473:pathology; D013045:Species Specificity; D013188:Sri Lanka
chemical_list: D017931:DNA Primers; D016054:DNA, Protozoan