Diagnosis and Characterization of Leishmania Species in Giemsa-Stained slides by PCR-RFLP
Background: Direct identification of Leishmania species in Giemsa-stained slides without parasite culturing in the areas where multiple species exist, is very helpful. This study was designed to isolate Leishmani spp. from Giemsa-stained smears and to characterize them by PCR technique. Methods: A...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Tehran University of Medical Sciences
2008-03-01
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| Series: | Iranian Journal of Public Health |
| Subjects: | |
| Online Access: | https://ijph.tums.ac.ir/index.php/ijph/article/view/2073 |
| Summary: | Background: Direct identification of Leishmania species in Giemsa-stained slides without parasite culturing in the areas where multiple species exist, is very helpful. This study was designed to isolate Leishmani spp. from Giemsa-stained smears and to characterize them by PCR technique.
Methods: A total of 48 Giemsa-Stained slides from confirmed cases of leishmaniasis were examined under a light microscope at×1000 and classified based on grading of Leishmania parasites. DNA from each slide was extracted separately and subjected to PCR. The ribosomal internal transcribed spacer 1 (ITS1) was amplified with specific primers and the PCR products were digested with a restriction enzyme (HaeIII).
Results: Of the 48 microscopy-positive slides, 43(89.6%) were positive by PCR-RFLP and Leishmania species were identified. A statistically significant difference was observed between the both methods (P< 0.05) and also a concordance was found between microscopy and PCR-RFLP (k= 0.55).
Conclusion: PCR-RFLP seems to be an effective method to identify Leishmania species from Giemsa-stained smears which have been collected from both infected humans and animal reservoir hosts in Iran. |
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| ISSN: | 2251-6085 2251-6093 |