Direct diagnosis of cutaneous leishmaniasis of skin lesion specimens by PCR and evaluate the sensitivity of testing methods in Wasit Province

Altamemy,; Aakool, A.K.

	Direct diagnosis of cutaneous leishmaniasis of skin lesion specimens by PCR and evaluate the sensitivity of testing methods in Wasit Province
Journal of College of Education / Wasit
Article 1, Volume 1, Issue 20, 2018, Pages 543-558
Authors
; A.K. Aakool
Abstract
The study had been conducted to diagnose the parasite that cause cutaneous leishmaniasis by using two specific primer , ten samples from cutaneous lesions ( expected to be cutaneous leishmaniasis) were collected from different ages males and females , some of sample spotted onto filter disks were punched out with a paper puncher (two disks per sample) to be used in the polymerase chain reaction (PCR) by using two different primers, the first was spliced leader mini-exon polymerase chain reaction (SLME PCR) used to idetify Leishmania parasites genus according to molecular weight, there are four samples with DNA bands molecular weight 89 bp and the other six samples with 100 bp. The sensitivity of this test was 100%. The second primer ( B6) is a specific primer that used to identify parasite species. Six samples of ten was positive and the molecular weight of DNA bands ranging from 359 – 360 bp, this refer to L. tropica, so the other four samples may be L. major because only two species of cutaneous leishmaniasis regionally prevalence in Iraq. Microscopic slides were prepared from the cutaneous lesion, stained by giemsa stain, the slides showed only eight cases positive by detecting the amastigotes in macrophages, so the sensitivity of this test was 80%. Samples of cutaneous lesion also cultured in a semi-solid culture medium and incubated at 26˚c , it had been noticed that there is Leishmania parasite in promastigote phase in eight of ten samples, the sensitivity of this test was 80%.

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