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Behbood  2011 

Characterization of Leishmania isolated from Kala- azar infected patients in Kermanshah using PCR

Keywords: visceral leishmaniasis , Kala- azar , L. infantum , PCR

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Abstract:

Background: Visceral leishmaniasis (VL) or Kala-azar is an important parasitic disease caused by protozoan parasites of the genus Leishmania including L. donovani and L. infantum. Some evidences show that VL is present in some areas of Kermanshah province and this study aimed to characterize the causative agent of VL in this region.Methods: Bone marrow sample was obtained from 9 VL patients from Pediatric Hospital in Kermanshah. DNA was extracted from the microscopic slides and was checked by specific PCR to find out the species of the parasite. To do that, a segment of mini circle kinetoplast DNA was amplified, using LINR4 and LIN17 primers. All samples produced a 720 bp band in PCR reaction. Products of PCR were evaluated by electrophoresis using 1.5% gel agarose.Results: Anti Leishmania antibody was detected in all patients except one case by IFAT. Examination of bone marrow smears demonstrated numerous amastigotes of Leishmania (Leishman bodies) in the samples. The isolates were compared with reference strains and revealed that all isolates were L. infantum.Conclusions: Our findings showed that L. infantum is the most common VL in Kermanshah and one-step PCR reaction may be suitable method to detect species' characterization. Further study is necessary to explore other aspects of VL including seroprevalence and parasite reservoirs in this region.

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