Characterization of ITS1 gene of Leishmania infantum isolated from Iraqi patients with visceral leishmaniasis by PCR- RFLP and sequencing methods. | ||||
Delta Journal of Science | ||||
Article 7, Volume 36, Issue 3, December 2013, Page 179-185 PDF (1.17 MB) | ||||
Document Type: Research and Reference | ||||
DOI: 10.21608/djs.2013.139772 | ||||
View on SCiNiTO | ||||
Authors | ||||
Ilham A. Majeed1; Fatemeh Gaffarifar2; Nada Al-Bashier3 | ||||
1Medical Laboratory Dept. College of Health & Medical Technology/ Baghdad/ Iraq. | ||||
2Parasitology and Entomology Dept., Faculty of Medical Sciences, Tarbiat Modares University, | ||||
3Medical Research Center/ College of Medicine- Al-Nehrain University/ Baghdad/ Iraq. | ||||
Abstract | ||||
For the best of our knowledge, there is no information about molecular characterization of Iraqi isolates of visceral leishmaniasis, the present work aimed to characterize three different Iraqi isolates of Leishmania infantum by polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) and sequencing methods. Three isolates from bone marrow of Iraqi patients infected with kala azar were used in this study. The isolates were already diagnosed by isoenzyme as Leishmania infantum. Patients were inhabiting different parts of Baghdad. The samples after microscopic examination were cultured on modified NNN media. Then DNA was extracted for amplifying ITS1 (internal transcribed spacer 1) gene by PCR. Identification of samples was studied using RFLP (digestion with Apo1 restriction enzyme) and sequencing of PCR products. The PCR of all samples showed a band under about 500 bp. The results by using PCR-RFLP method showed no restriction with digestion with Apo1 restriction enzyme. The results of sequencing showed differences with all separated gene from Leishmania infantum in the gene bank. In this study we found that the sequences of ITS1 gene of Leishmania infantum separated from Iraqi patients are different from other samples, as there is no similarity with Leishmania infantum (MHOM/TN/80/IPI1). The more similarity is with the Iranian isolate of Leishmania infantum (MCAN/IR/97/LON) with 41%, whereas the similarity with Crithidia luciliae internal transcribed spacer 1, ITS1 is 96%. | ||||
Keywords | ||||
Visceral leishmaniasis; PCR-RFLP; sequencing; Iraq | ||||
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