Genotyping of Hydatid Cyst Isolated from Human and Domestic Animals in Ilam Province, Western Iran Using PCR-RFLP
-
M Dousti
,
J Abdi
,
S Bakhtiyari
,
M Mohebali
,
Sh Mirhendi
,
Mb Rokni
Abstract
Background: Hydatidosis or cystic hydatid disease is one of the most important diseases in human and animals. Identification of strains is important for improvement of control and prevention of dis-ease. The aim of this study was to determine the strains isolated from human and domestic animals in Ilam Province, Iran, using PCR-RFLP method.
Methods: Respectively, 30 and 4 animal and human hydatid cysts were collected from different slaughterhouses and hospitals of the province. Protoscolices were separated and their DNA genome was extracted by extraction kit. rDNA-ITS1 of each isolated samples was duplicated by BD1(Forward) and 4s (Reverse) Primers. PCR products were studied by electrophoresis and then were digested using TaqI, HpaII, RsaI and AluI restriction enzymes. RFLP products were studied using electrophoresis on 1% agar gel.
Result: A fragment of 1000bp was produced from amplification of rDNA-ITS1 of protoscolices using PCR method. After digestion of PCR product by AluI enzyme, 200bp and 800bp, by RsaI, 655bp and 345bp and by HpaII 700bp and 300bp sizes were obtained. TaqI enzyme had no change in fragment size and it remained 1000bp. Considering the method, Ilam strains was specified as E. granulosus sensu stricto (G1-G3).
Conclusions: Although sheep strain (G1) is dominated in human and different animal in Iran and the world, but more efforts should be done to clarify the true genotype of Ilam strains specified as E. granulosus sensu stricto (G1-G3).
Budke CM, Torgerson PR. Global Socioeconomic Impact of Cystic Echinoco-ccosis. Emerging Infectious Diseases. 2006;12(2): 296-303.
Abdi J, Kazemi B, Karimfar MH, Rokni MB. Evaluation of rabbit antibody response against 8 and 16 kDa recombinant subunits of antigen B from Echinococcus granulosus. Asian Pacific J Trop Med. 2012;5(5):355-7.
Rokni MB. Echinococcosis /hydatidosis in Iran. Iranian J Parasitol. 2009;4(1):1-16.
Heidari Z, Mohebali M, Zarei Z, Aryayipour M, Eshraghian MR, Kia EB, et al. Ser-oepidemiological study of human hydatidosis in Meshkinshahr district, ardabil province, Iran. Iranian J Parasitol. 2011;6(3): 19-25.
Rostami Nejad M, Nazemalhosseini Mojarad E, Nochi Z, Fasihi Harandi M, Cheraghipour K, Mowlavi GR, et al. Echinococcus granulosus strain differentiation in Iran based on sequence heterogeneity in the mitochondrial 12S rRNA gene. J Helminthol. 2008;82(04): 343-7.
Mehrabani D, Oryan A, Sadjjadi SM. Prevalence of Echinococcus granulosus infection in stray dogs and herbivores in Shiraz, Iran. Vet Parasitol. 1999;86(3):217-20.
Maleky F, Moradkhan M. Echinococcosis in the stray dogs of Tehran, Iran. Ann Trop Med Parasitol. 2000;94(4):329-31.
Eslami A, Hosseini SH. Echinococcus granulosus infection of farm dogs in Iran. Parasitol Res. 1998;84(3):205-7.
Ahmadi NA. Hydatidosis in camels (Camelus dromedarius) and their potential role in the epidemiology of Echinococcus granulosus in Iran. J Helminthol. 2005 Jun;79(2):119-25.
Aflaki A, Ghaffarifar F, Dalimi Asl A. Seroepidemiological survey of hydatidosis by Dot- ELISA in Ilam province. Journal of Medical Science of Modarres. 2005;8(1):1-6.
Abdi J, Kazemi B, Mohebali M, Bandehpour M, Rahimi MT, Rokni MB. Gene cloning, expression and serological evaluation of the 12-kDa antigen-B subunit from Echinococcus granulosus. Ann Trop Med Parasitol. 2010;104(5):399-407.
Bowles J, Blair D, McManus DP. Genetic variants within the genus Echinococcus identified by mitochondrial DNA sequencing. Mol Biochem Parasitol. 1992;54(2):165-73.
McManus DP RA. Genetic heterogeneity within Echinococcus granulosus: isolates from different hosts and geographical areas characterized with DNA probes. Parasitology. 1989;99 (1):17-29.
Abdi J, Kazemi B, Haniloo A, Mohebali M, Mahmoudi M, Rezaei S, et al. Serological evaluation of EgAgB16 kDa, a recombinant antigen from Echinococcus granulosus for diagnosis of human hydatidosis. Iranian J Parasitol. 2010;5(3):1-10.
Bowles J, McManus DP. Rapid disc-rimination of Echinococcus species and strains using a polymerase chain reaction-based RFLP method. Mol Biochem Parasitol. 1993; 57(2): 231-9.
Hop M, Bowles J, McManus DP. A reconsideration of the Echinococcus granulosus strain in Australoa following RFLP analysis of cystic material. Int J Parasitol. 1997;21(4):471-5.
Yoosefi H, Hashemzadeh M, Aliyari Z. Molecular study of hydatid cyst (sheep strain) in Chaahrmohal va Bakhteyari by PCR-RFLP. J Shahrkord Uni. 2007;9(2):28-33.
Shahnazi M. Molecular characterization of human and animal Echinococcus granulosus isolates in Isfahan, Iran. Acta Trop. 2011;117(1):47-50.
Zhang L, Eslami A, Hosseini SH, McManus DP. Indication of the presence of two distinct strains of Echinococcus granulosus in Iran by mitochondrial DNA markers. Am J Trop Med Hyg. 1998;59(1):171-4.
Yakhchali M, Mardani K. Study on strain variation of Echinococcus granulosus in domestic life cycle by amplification of nad-1 gene by PCR-RFLP. Iran J Vet. 2011;7(1):63-8.
Sadri A, Moshfe A, Doosti A. Characterizat-ion of Isolated Hydatid Cyst from Slaughtered Livestock in Yasuj Industrial Slaughterhouse by PCR-RFLP. J Yasuj Uni. 2012;17(3):143-52.
Pezeshki A, Akhlaghi L. Genotyping of Echinococcus granulosus from domestic animals and humans from Ardabil Province, northwest Iran. J Helminthol. 2012;16(2):1-5.
Sharbatkhori M, Fasihi Harandi M. Sequence analysis of cox1 and nad1 genes in Echinococcus granulosus G3 genotype in camels (Camelus dromedarius) from central Iran. Parasitol Res. 2011;108(3):521-7.
Varcasia A, Canu S, Kogkos A, Pipia AP, Scala A, Garippa G, et al. Molecular characterization of Echinococcus granulosus in sheep and goats of Peloponnesus, Greece. Parasitol Res. 2007 Sep;101(4):1135-9.
Mc Manus DP, Thompson RC. Molecular epidemiology of cystic echinococcosis. Parasitology. 2003;127: 37-51.
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| Issue | Vol 8 No 1 (2013) | |
| Section | Articles | |
| Keywords | ||
| Echinococcus granulosus Genotyping Iran Protoscolices Strain | ||
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