Molecular Diagnosis and Phylogenetic Analysis of 5.8s rDNA Gene of Cutaneous Leishmaniasis Species in Holy Karbala/Iraq Hamza Dhamiaa Maki1, Obayes Laith Hasan1, Zghair Fadhil Sami2 1Dept. of Microbiology, College of Medicine, Karbala University, Karbala, Iraq 2Dept. of Medical Laboratory, Babylon technical institute, Al-Furat Al-Awsat Technical University, Hilla, Iraq Online published on 6 April, 2019. Abstract Objective To identify Leishmania species in patients with cutaneous leishmaniasis (CL) in the Holy Karblala in Iraq from November 2017 to the end of January 2018. Method Among 100 patients were examined by dermatologists for CL at Al-Husseini Hospital with suspected stained slide smears of 92 patients showed positive result for Leishmania., The aspirated material was inoculated in Biphasic Medium which used for maintaining and cultivating Leishmania spp. polymerase chain reaction (PCR) protocols were used to amplify a region of the ribosomal DNA amplicon of Leishmania (5.8s rDNA gene). Sequencing of amplified products was done by Microgen company/USA through using by forward primer for 5.8s rDNA gene in sequencing reactions by Applied Bio system ABI Prism 377 DNA sequencer at Molecular biology department in Korea. Results Of 92 positive sample of Leishmania, 20 (21.7%) belonged to females and 72 (78.3%) belonged males. The PCR assays detected Leishmania major in all samples their ages ranged from (10–19 years) 6(6.5%), while ages ranged from (20–29 years) 11(11.9%), and ages ranged from (30–39 years) 17(18.5%) while the age (40–49) 37 (40.2%) and (50–59) 21 (22.8). The occupation in kids 3(3.2%), students 11(11.9%), solders 34(36.9%), workers 20 (21.7%), Employees 9 (9.8) and house wives 15(16.3%). Most of the lesions were found on the exposed areas of the body: face 25 (27.2%), hands 44(47.8%), Legs 11(12%), and ears 12(13%) Conclusion Cutaneous leishmaniasis with diverse clinical manifestation, is prevalent in karbala city caused by L.major and all of isolates is similar in 5.8s rDNA gene sequence closest with Iranian isolate genetically and distant for chinese isolate. Top Keywords Cutaneous leishmaniasis, Leishmania major, Molecular diagnosis, phylogeny, 5.8S rDNA Gene. Top |