Journal of Pure and Applied MicrobiologyVol. 8 No. 6

Molecular Diagnosis and Characterization of Leishmania Species from Clinical Specimens in Kashan, Iran 2012 - 2013

Baharak Ghorbanzadeh1, Sima Rasti1*, Farnaz Kheirandish2, Ahmad Piroozmand3, Gholamabbas Mousavi4 and Bathol Abani5

1Department of Parasitology, Faculty of Medicine, Kashan University of Medical Sciences, Kashan Iran. 2Department of Medical Parasitology and Mycology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran. 3Department of Microbiology and Immunology, Faculty of Medicine, Kashan University of Medical Sciences. Kashan Iran. 4Department of Statistics and Public Health, Faculty of Health, Kashan University of Medical Sciences, Kashan. Iran. 5Zidi Health Care and Treatment Center, Kashan University of Medical Sciences. Kashan Iran.

Received on 02 August 2014 and accepted on 01 September 2014

 

ABSTRACT

Cutaneous Leishmaniasis is one of the health problems in Mediterranean regions including Iran. Considering the increase in the incidence of disease in Kashan, this study was designed to identify the Leishmania species in cutaneous leishmaniasis by PCR using primers of kinetoplastic DNA for treatment and appropriate measure for controlling of the disease. 130 patients suspected for cutaneous leishmaniosis referred to two health care centers of Kashan from 2012 to 2013 were examined. The demographic information as well as signs of the disease were recorded. The diagnosis of infection was based on observation of amastigotes within the smear of, serosity of the wound, after dying by Giemsa. Then, Leishmania species was identified following the extraction of DNA from serosity, and PCR with variable region of the KDNA. Expected PCR products of L. major and L. tropica were 650 bp and 760 bp respectively. The data were analyzed by SPSS using X2 and Kappa (K). Overall of 130 specimens, 87(66.9%) cases and 98 (75.4 %) cases were positive for microscopy and PCR methods respectively. In PCR assay 70 (71.4 %) and 26 (26.6%) of the samples were identified as L. tropica and L. major respectively, and 2 (2 %) cases were detected as mix. The majority of the L.tropica or L. major had 1 to 2 wounds, but 11.5 % of L. major positive cases had more than 7 wounds respectively. Based on the results of this study, both species of Leishmania were present in Kashan, Therefore, it was suggested that careful preventive measure be taken in rural and urban parts. In addition, KDNA-PCR assay using serosity for accurate and quick diagnosis of cutaneous leishmaniasis is recommended.

Keywords : Cutaneous Leishmaniasis, PCR, KDNA, Molecular Characterization.