Author/Authors :
HOSSEINI FARASH, Bibi Razieh Dept. of Medical Parasitology and Mycology - School of Public Health - Tehran University of Medical Sciences, Tehran, Iran , MOHEBALI, Mehdi Center for Research of Endemic Parasites of Iran - Tehran University of Medical Sciences, Tehran, Iran , KAZEMI, Bahram Dept. of Biotechnology - School of Medicine - Shahid Beheshti University of Medical Sciences, Tehran, Iran , HAJJARAN, Homa Dept. of Medical Parasitology and Mycology - School of Public Health - Tehran University of Medical Sciences, Tehran, Iran , AKHOUNDI, Behnaz Dept. of Medical Parasitology and Mycology - School of Public Health - Tehran University of Medical Sciences, Tehran, Iran , RAOOFIAN, Reza Legal Medicine Research Center - Iranian Legal Medicine Organization, Tehran, Iran , FATA, Abdolmajid Dept. of Medical Parasitology and Mycology - School of Medicine - Mashhad University of Medical Sciences, Mashhad, Iran , MOJARRAD, Majid Dept. of Medical Parasitology and Mycology - School of Medicine - Mashhad University of Medical Sciences, Mashhad, Iran , SHARIFI-YAZDI, Mohammad Kazem Zoonosis Research Center - Tehran University of Medical Sciences, Tehran, Iran
Abstract :
Background: Visceral leishmaniasis (VL) caused by Leishmania infantum is the most severe form of leishmaniasis in Iran, which causes a high mortality rate in the case of inaccurate diagnosis and treatment. This study aimed to clone of K26 gene from Iranian strain of L. infantum and register the sequencing results in Genbank to facilitate the preparation a new K26 antigen for the detection of L. infantum infection.
Methods: L. infantum was obtained from an infected domestic dog in Meshkin-Shahr area from northwestern Iran in 2015. Canine visceral leishmaniasis was confirmed by direct agglutination test (DAT), rK39 dipstick and parasitological methods. L. infantum was confirmed by N-acetyl glucosamine -1-phosphate transferase (nagt)–PCR and its sequencing. The band of interest for k26 form Iranian strain of L. infantum was purified by gel extraction kit after PCR amplifica-tion and then ligated into pBluescript II SK (+) and pET-32a (+), respectively. The sequences of recombinant plas-mids were analyzed and submitted to Genbank.
Results: The submission of rk26 nucleotide sequence was performed to the GeneBank/NCBI Data Base under ac-cession number KY212883. The related gene was showed a homology about 99% to L. chagasi and L. infantum k26 gene, while the level of homology in comparison with different strains of L. donovani ranged from 84-94%.
Conclusion: The successful rk26 cloning into an expression vector performed in this study could help to produce a new recombinant antigen for serodiagnosis of VL especially in areas where L. infantum is the main causative agent.
