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Background: Leishmaniasis
is a parasitic infectious disease which causes skin sores. There is no
effective laboratory screening tests for leishmaniasis. Some diagnostic
techniques exist that allow parasite detection and species identification by
special culture and microscopy, biochemical (Isoenzymes), immunologic
(immunoassays), and molecular (PCR) approaches. Specific major objectives of
this study was to genotyping of Leishmania species in Bam and Shiraz city.
Methods: A total of 83
samples of Leishmania were collected from patients clinically suspected of cutaneous
leishmaniasis. The geographic distributions of the samples were 55 samples from
Bam and 28 from Shiraz city. For this propose samples of skin and bloods were
blotted on filter paper. Genomic DNA extracted with a Genomic DNA extraction
kit (AccuPrep, BIONEER). Aliquots of extracted DNA were kept at -20°C.
region of ITS1 amplified with the published Leishmania-specific primers. 15-20mL of these amplicons,
containing the amplified ITS1 region, was digested for 2h with HaeIII.
Results: All 55 samples from
Bam were considered as L. tropica and the positive samples from Shiraz
considered as L. tropica and just one sample was L. major which was belonged to
a patient had previously traveled to Isfahan and Khuzestan.
Conclusion: In
the current study a PCR technique was employed for amplification of Leishmania DNA
directly in biological materials. Characterization of genus of Leishmania using
RFLP-PCR method is too sensitive and too rapid, and there is no need for culturing
the parasite for diagnosis.
