Tracking of Coetaneous Leishmaniasis by Parasitological, Molecular and Biochemical Analysis

Abstract

Background: In areas of endemicity without sufficient laboratory infrastructure, identification of Leishmaniaparasites is useful for control and preventive plan and very important since species differentiation is instrumental in selection of optimal therapy and treatment regimens. objectives: The present study was performed to identify the species and strain of Leishmaniaparasites isolated of different endemic areas. Methodology: In a consecutive series of 126 patients referred for a suspected CL lesion during October 2010 to December 2012 in five Iraqi provinces, direct smear and culture followed by molecular and biochemical analysis were done using nested-PCR and cellulose acetate electrophoresis. Results: Direct smears revealed that 90 (70%) patients gave positive results under light microscope. Out of 126 cases, 83 (467) gave positive growth in modified NNN medium and RPMI-1640 with fetal bovine serum followed by sub-culture in drosophila Schneider’s media. Out of 83 positive growth culture, only 52 specimens were studied by nested - PCR .It was found that 45 (89.5%) cases in the generation of a 560 bp DNA and 7(13.4%) patients displayed a fragment of 750 bp, corresponding to L. major and L. tropica, respectively. Cellulose acetate electrophoresis (CAE) was performed by 4 enzyme systems (glucose phosphate isomerize, leucilphosphate, mannose phosphate isomerize and 6 phosphogluconate dehydrogenase). The results showed that L. Major(LV39) isolate were in 20 mass cultivated culture Conclusion: Nested-PCR and cellulose acetate electrophoresis was an ideal method for discrimination of Leis mania. spp. and variants. Both L. major and L. tropica were the causative agents of cutaneous Leishmaniasis but L. major was the main species in study area. Recommendations: As a result of the observations made during the whole study period, nested- PCR is very suitable method, which can be used as a basis for future applications. Opening number of research centers situated in the country offer DNA applications for so-called “parasite tracking.” Such an application is probably more relevant for epidemiological purposes than for diagnosis. (e.g., in outbreak investigations or tracking drug-resistant parasite strains). After collection in the field, samples can be simply transferred to such a center for further analysis. More experimental studies for effective vaccine are developed for Leishmaniasis.