Molecular Identification and Differentiation of Fasciola Isolates Using PCR- RFLP Method Based on Internal Transcribed Spacer (ITS1, 5.8S rDNA, ITS2)
Background: In this study, we used both ITS1 and ITS2 for molecular identification of Fasciola species.Methods: The region between 18S and 28S of ribosomal DNA was used in PCR-RFLP method for molecular identification of Fasciola species. Ninety trematodes of Fasciola were collected during abattoir i...
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Tehran University of Medical Sciences
2011-09-01
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doaj-72fcde0788104ff3930d546301a42ce42021-03-02T11:00:07ZengTehran University of Medical SciencesIranian Journal of Parasitology1735-70202008-238X2011-09-01633542Molecular Identification and Differentiation of Fasciola Isolates Using PCR- RFLP Method Based on Internal Transcribed Spacer (ITS1, 5.8S rDNA, ITS2) M Forouzandeh-MoghadamMB RokniA DalimiM Mahami-OskoueiBackground: In this study, we used both ITS1 and ITS2 for molecular identification of Fasciola species.Methods: The region between 18S and 28S of ribosomal DNA was used in PCR-RFLP method for molecular identification of Fasciola species. Ninety trematodes of Fasciola were collected during abattoir inspection from livers of naturally infected sheep and cattle from Khorasan, East Azerbaijan, and Fars provinces in Iran. After DNA extraction, PCR was performed to amplify region ITS1, 5.8S rDNA, ITS2. To select a suitable restriction enzyme, we sequenced and ana-lyzed the PCR products of F. hepatica and F. gigantica samples from sheep and cattle. Tsp509I fast digest restriction enzyme was selected for RFLP method that caused the separation specifi-cally of Fasciola species. Results: The fragment approximately 1000bp in all of the Fasciola samples was amplified and then digested with the Tsp509I restriction endonuclease. Seventy F. hepatica and 20 F. gigantica were identified of total 90 Fasciola isolates.Conclusion: The new PCR-RFLP assay using Tsp509I restriction enzyme provides a simple, practical, fast, low cost, and reliable method for identification and differentiation of Fasciola isolates.http://journals.tums.ac.ir/upload_files/pdf/19098.pdfFasciola HepaticaFasciola GiganticaITS15.8S rDNAITS2PCR-RFLPIran |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
M Forouzandeh-Moghadam MB Rokni A Dalimi M Mahami-Oskouei |
spellingShingle |
M Forouzandeh-Moghadam MB Rokni A Dalimi M Mahami-Oskouei Molecular Identification and Differentiation of Fasciola Isolates Using PCR- RFLP Method Based on Internal Transcribed Spacer (ITS1, 5.8S rDNA, ITS2) Iranian Journal of Parasitology Fasciola Hepatica Fasciola Gigantica ITS1 5.8S rDNA ITS2 PCR-RFLP Iran |
author_facet |
M Forouzandeh-Moghadam MB Rokni A Dalimi M Mahami-Oskouei |
author_sort |
M Forouzandeh-Moghadam |
title |
Molecular Identification and Differentiation of Fasciola Isolates Using PCR- RFLP Method Based on Internal Transcribed Spacer (ITS1, 5.8S rDNA, ITS2) |
title_short |
Molecular Identification and Differentiation of Fasciola Isolates Using PCR- RFLP Method Based on Internal Transcribed Spacer (ITS1, 5.8S rDNA, ITS2) |
title_full |
Molecular Identification and Differentiation of Fasciola Isolates Using PCR- RFLP Method Based on Internal Transcribed Spacer (ITS1, 5.8S rDNA, ITS2) |
title_fullStr |
Molecular Identification and Differentiation of Fasciola Isolates Using PCR- RFLP Method Based on Internal Transcribed Spacer (ITS1, 5.8S rDNA, ITS2) |
title_full_unstemmed |
Molecular Identification and Differentiation of Fasciola Isolates Using PCR- RFLP Method Based on Internal Transcribed Spacer (ITS1, 5.8S rDNA, ITS2) |
title_sort |
molecular identification and differentiation of fasciola isolates using pcr- rflp method based on internal transcribed spacer (its1, 5.8s rdna, its2) |
publisher |
Tehran University of Medical Sciences |
series |
Iranian Journal of Parasitology |
issn |
1735-7020 2008-238X |
publishDate |
2011-09-01 |
description |
Background: In this study, we used both ITS1 and ITS2 for molecular identification of Fasciola species.Methods: The region between 18S and 28S of ribosomal DNA was used in PCR-RFLP method for molecular identification of Fasciola species. Ninety trematodes of Fasciola were collected during abattoir inspection from livers of naturally infected sheep and cattle from Khorasan, East Azerbaijan, and Fars provinces in Iran. After DNA extraction, PCR was performed to amplify region ITS1, 5.8S rDNA, ITS2. To select a suitable restriction enzyme, we sequenced and ana-lyzed the PCR products of F. hepatica and F. gigantica samples from sheep and cattle. Tsp509I fast digest restriction enzyme was selected for RFLP method that caused the separation specifi-cally of Fasciola species. Results: The fragment approximately 1000bp in all of the Fasciola samples was amplified and then digested with the Tsp509I restriction endonuclease. Seventy F. hepatica and 20 F. gigantica were identified of total 90 Fasciola isolates.Conclusion: The new PCR-RFLP assay using Tsp509I restriction enzyme provides a simple, practical, fast, low cost, and reliable method for identification and differentiation of Fasciola isolates. |
topic |
Fasciola Hepatica Fasciola Gigantica ITS1 5.8S rDNA ITS2 PCR-RFLP Iran |
url |
http://journals.tums.ac.ir/upload_files/pdf/19098.pdf |
work_keys_str_mv |
AT mforouzandehmoghadam molecularidentificationanddifferentiationoffasciolaisolatesusingpcrrflpmethodbasedoninternaltranscribedspacerits158srdnaits2 AT mbrokni molecularidentificationanddifferentiationoffasciolaisolatesusingpcrrflpmethodbasedoninternaltranscribedspacerits158srdnaits2 AT adalimi molecularidentificationanddifferentiationoffasciolaisolatesusingpcrrflpmethodbasedoninternaltranscribedspacerits158srdnaits2 AT mmahamioskouei molecularidentificationanddifferentiationoffasciolaisolatesusingpcrrflpmethodbasedoninternaltranscribedspacerits158srdnaits2 |
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