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. 2012;7(4):8-16.

Molecular Characterization of Echinococcus granulosus from Hydatid Cysts Isolated from Human and Animals in Golestan Province, North of Iran

Sh Gholami  1 , M Sosari, M Fakhar, M Sharif, A Daryani, Mb Hashemi, M Vahadi
Affiliations

Molecular Characterization of Echinococcus granulosus from Hydatid Cysts Isolated from Human and Animals in Golestan Province, North of Iran

Sh Gholami et al. Iran J Parasitol. 2012.

Abstract

Background: The aim of the present study was to determine the molecular characteristics of Echinococcus granulosus from paraffin-embedded tissues of hydatid cysts isolated from human and protoscoleces of hydatid cysts from sheep, cattle and camel isolates using PCR- RFLP of ITS1- rDNA analysis in Golestan Province, northern Iran.

Methods: E. granulosus isolates from human patients infected with hydatid cyst and protoscoleces from hydatid cysts of sheep, cattle and camel isolates were collected from different hospitals and the abattoir throughout the Golestan Province. In all, 60 E. granulosus genomic DNA were extracted and examined by PCR - ITS1 of rDNA and amplified using BD1 / 4S and EGF1 / EGR2 primers, followed by RFLP using Alu1, Msp1 and TaqI restriction enzymes.

Results: The PCR-ITS1 products obtained from sheep, cattle and human isolates were similar to sheep strain (1000 bp and 391 bp). Majority of the camel samples yielded 295 bp DNA bands. RFLP -ITS1 of E. granulosus with Taq1 in human, sheep and cattle isolates showed similar patterns in the number and size of DNA. RFLP methods in camel isolates showed a different genotype, using Taq1, whereas no DNA bands were observed using Alu1 in camel and human isolates. Therefore, two clearly distinguishable banding patterns of E. granulosus were obtained with the three enzymes, which separating human, sheep and cattle isolates from the camel origin.

Conclusion: The results indicate the possible of transmission of the G1 and G6 genotypes of E. granulosus between livestock animals and human in Golestan Province.

Keywords: Echinococcus granulosus; Hydatid cysts; Iran; Molecular Characterization.

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Figures

Fig. 1
Fig. 1
PCR amplified ITS1 fragments from various isolates of E. granulosus from Golestan, Iran SH: Sheep (size, 1000 bp and 900 bp)/ HU: Human (size, 1000 bp and 391) /CA: Cattle liver (size, 1391bp)/CM: Camel (size, 295 bp)/ N: Negative control (without DNA template)/ L: DNA lader
Fig. 2
Fig. 2
RFLP fragments of various isolates of E. granulosus from Golestan, Iran. PCR amplified ITS1 products were digested with Taq1: SH1: Sheep (size, 334,137 bp) SH2: Sheep (size, 650,250,150 bp) HU1: Human (size, 281,110) HU2: Human (size, 650,250,150 bp) CA: Cattle (size, 281, 110 bp) CM: Camel (size, 281,110) L: DNA lader
Fig. 3
Fig. 3
RFLP fragments of various isolates of E. granulosus from Golestan, Iran. PCR amplified ITS1 products were digested with Alu1: CA1: Cattle (391bp) undigested CA2: Cattle (334,137 bp) CA3: Cattle (500,400,100 bp) CM: Camel (295 bp) undigested SH1: Sheep (500 bp) undigested SH2: Sheep (700 bp, 300 bp) L: DNA lader
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