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Polymerase chain reaction detection of LeishmaniaDNA in skin biopsy samples in Sri Lanka where the causative agent of cutaneous leishmaniasis is Leishmania donovani
Ranasinghe, Shalindra; Wickremasinghe, Renu; Hulangamuwa, Sanjeeva; Sirimanna, Ganga; Opathella, Nandimithra; Maingon, Rhaiza DC; Chandrasekharan, Vishvanath.
  • Ranasinghe, Shalindra; University of Sri Jayewardenepura. Faculty of Medical Sciences. Department of Parasitology. Nugegoda. LK
  • Wickremasinghe, Renu; University of Sri Jayewardenepura. Faculty of Medical Sciences. Department of Parasitology. Nugegoda. LK
  • Hulangamuwa, Sanjeeva; University of Sri Jayewardenepura. Faculty of Medical Sciences. Department of Parasitology. Nugegoda. LK
  • Sirimanna, Ganga; University of Sri Jayewardenepura. Faculty of Medical Sciences. Department of Parasitology. Nugegoda. LK
  • Opathella, Nandimithra; University of Sri Jayewardenepura. Faculty of Medical Sciences. Department of Parasitology. Nugegoda. LK
  • Maingon, Rhaiza DC; University of Sri Jayewardenepura. Faculty of Medical Sciences. Department of Parasitology. Nugegoda. LK
  • Chandrasekharan, Vishvanath; University of Sri Jayewardenepura. Faculty of Medical Sciences. Department of Parasitology. Nugegoda. LK
Mem. Inst. Oswaldo Cruz ; 110(8): 1017-1023, Dec. 2015. tab, graf
Article in English | LILACS | ID: lil-769836
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.
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Full text: Available Index: LILACS (Americas) Main subject: Skin / Leishmania donovani / Polymerase Chain Reaction / DNA, Protozoan / Leishmaniasis, Cutaneous Type of study: Diagnostic study / Prognostic study Limits: Humans Country/Region as subject: Asia Language: English Journal: Mem. Inst. Oswaldo Cruz Journal subject: Tropical Medicine / Parasitology Year: 2015 Type: Article / Project document Affiliation country: Sri Lanka Institution/Affiliation country: University of Sri Jayewardenepura/LK

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Full text: Available Index: LILACS (Americas) Main subject: Skin / Leishmania donovani / Polymerase Chain Reaction / DNA, Protozoan / Leishmaniasis, Cutaneous Type of study: Diagnostic study / Prognostic study Limits: Humans Country/Region as subject: Asia Language: English Journal: Mem. Inst. Oswaldo Cruz Journal subject: Tropical Medicine / Parasitology Year: 2015 Type: Article / Project document Affiliation country: Sri Lanka Institution/Affiliation country: University of Sri Jayewardenepura/LK