Identification of Leishmania species by high-resolution melting analysis in newly emerged foci in Sabzevar, northeast of
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ORIGINAL ARTICLE
Identification of Leishmania species by high-resolution melting analysis in newly emerged foci in Sabzevar, northeast of Iran Mohammad Javad Namazi1,2 • Ehsan Javaheri1,3 • Hadi Atabati1 • MohammadShafi Mojadadi1
Received: 23 April 2020 / Accepted: 14 October 2020 Ó Indian Society for Parasitology 2020
Abstract Cutaneous leishmaniasis (CL) is a zoonotic disease with 1 to 1.5 million annual incidences. Microscopic examination of the Giemsa stained slides is the most common diagnostic method for CL. However, this method cannot distinguish leishmania species. Hence the present study was conducted to identify leishmania species by high-resolution melting (HRM) analysis in the newly emerged foci of CL in Sabzevar, northeast of Iran. In this cross-sectional study, fifty patients with suspicious cutaneous lesions referring to the designated health center for diagnosis and treatment of CL in Sabzevar during 2017–2018, were recruited. All collected samples and prepared slides were stained for microscopic examination and then undergone HRM real-time PCR (HRM-PCR) assay to identify species of Leishmania parasites. The results of HRM-PCR technique showed that Leishmania major (L. major) was the dominant causative parasite in the newly emerged foci whereas L. tropica (L. tropica) was positive only in two patients. This was the first time that 7SL RNA-HRM-PCR assay was performed to precisely identify leishmania parasites in the northeast of Iran. We proved the newfound foci in which both L. major and L. Mohammad Javad Namazi and Ehsan Javaheri both authors contributed equally to this work. & Mohammad-Shafi Mojadadi [email protected] 1
Leishmaniasis Research Center, Sabzevar University of Medical Sciences, Sabzevar, Iran
2
Institute of Infection, Immunity and Inflammation. College of Medical, Veterinary and Life Science, University of Glasgow, Glasgow, UK
3
Cellular and Molecular Research Center, Sabzevar University of Medical Sciences, Sabzevar, Iran
tropica were present. In contrast to the recent studies which identified only L. major in the region, we showed that L. tropica was still present. Keywords High resolution melting analysis Real-time polymerase chain reaction Leishmaniasis Leishmania tropica Leishmania major
Introduction Leishmaniasis is a collective name for historically known diseases in human and animals with three different clinical characteristics including cutaneous, muco-cutaneous and potentially fatal visceral types (WHO 2010). Cutaneous leishmaniasis (CL) has two divergent clinical manifestations, rural (wet) and urban (dry). They are so called zoonotic CL (ZCL) and anthroponotic CL (ACL) due to different reservoirs and clinical aspects. Although the pathogenicity of both ACL and ZCL are mainly similar, differences are shorter period, more intense and expanded lesions with faster lesion healing in ZCL compared to ACL (Sergiev et al. 2018). Worldwide population at risk of leishmaniasis is estimated to be 350 million in 88 countries, annual prevalence is twelve million,
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