Molecular characterization of the human and sheep hydatid cyst strains in Chaharmahal va Bakhtiari province of Iran using restriction fragment length plolymorphism (PCR RFLP)
M.Moazeni, S.Taghipoor, M.Abolhasani, M.Hashemzadeh, E.Zarean, Hossein Yousofi Darani
1Dept of Sergery,Kashani hospital,Faculty ofMedicine, ShahrekordUniversity ofMedicalSciences, Shahrekord, (IRAN)
2Cellular andmolecular research center, ShahrekordUniversity ofMedical Sciences, Shahrekord, (IRAN)
3Dept of Parasitology&Mycology, Faculty ofMedicine, IsfahanUniversity ofMedical Sciences, Isfahan, (IRAN)
Background: Hydatidosis caused by larval stage of Echinococcus granulosus is a cosmopolitan zoonotic infection. In endemic area for this disease there is considerable genetic variation among different isolates of the parasite. These variations may affect the epidemiology, pathology and control of the disease. In this work strain identification of hydatid cysts isolated fromhuman or sheep in Chaharmahal va Bakhtiari province of Iran has been investigated.
Method: Fertile sheep hydatid cysts were collected fromseveral abattoirs inChaharmahal vaBakhtiari province of Iran.Human isolateswere obtained at surgery from Kashani hospital in the same area. DNA was extracted from preserved protoscoleces and Nested PCR was performed on the extracted DNA samples, the rDNA-ITS fragment was amplified subsequently. Using 4 restriction enzymes include Rsa², HpaI I, Alu² and taq², PCR-RFLP procedure was performed on the PCR products.
Results: The size of PCR product in this research was 1000bp in both human and sheep isolates. UsingAlu² enzyme; three fragments of 100, 180 and 720bp in human isolates and two fragments of 800bp and 200bp in sheep isolateswere created. Rsa² also revealed three segments of 150, 180 and 670bp in human samples and two fragments of 655bp and 345bp in sheep samples. After using HpaII enzyme three segments with 120, 200 and 680bp length in human isolates and two fragments with 700bp and 300bp in sheep isolates were detected. Finally using Taq² enzyme no digestion was occurred on human or sheep samples.
Conclusion: The result of this investigation showed that human hydatid cyst strain in Chaharmahal & Bakhtiati province of Iran is different from sheep ones, so it is recommended to recognize DNA sequence in this human samples in future studies. ! 2013 Trade Science Inc. - INDIA
Hydatid cyst; RFLP; Strain; Human; Sheep.