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Evaluation of a Single PCR Assays on Cp5 Gene for Differentiation of Entamoeba histolytica and E. dispar
BACKGROUND: We examined a molecular method with a single-PCR for amplification of a part of CP5 gene enabling us to differentiate the pathogenic species, Entamoeba histolytica, from the non-pathogenic species, E. dispar. METHODS: We developed a single PCR method for this purpose. After investigation...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Tehran University of Medical Sciences
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3481692/ https://www.ncbi.nlm.nih.gov/pubmed/23113039 |
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author | Rostamighalehjaghi, S Jamali, R Rezaie, S Babaei, Z Hooshyar, H Rezaeian, M |
author_facet | Rostamighalehjaghi, S Jamali, R Rezaie, S Babaei, Z Hooshyar, H Rezaeian, M |
author_sort | Rostamighalehjaghi, S |
collection | PubMed |
description | BACKGROUND: We examined a molecular method with a single-PCR for amplification of a part of CP5 gene enabling us to differentiate the pathogenic species, Entamoeba histolytica, from the non-pathogenic species, E. dispar. METHODS: We developed a single PCR method for this purpose. After investigation of GenBank, primer pairs were designed from highly conserved regions of cysteine proteinase (CP5) gene. The primers were utilized in PCR using isolated genomic DNA template of E. histolytica and the PCR products were then sequenced. The same primer and method for PCR was used for isolated genomic DNA template of E. dispar. RESULTS: A fragment of about 950 bp was isolated in PCR by using DNA from E. histolytica, however, no banding pattern was produced by using the same primers for E. dispar. We characterized CP5 gene at molecular level in E. histolytica isolates from 22 positive; including 20 non-dysentery samples isolated from both cities as well as two dysentery samples isolated only from Tabriz. Nucleotide sequence comparison in gene data banks (NCBI, NIH) revealed significant homology with CP5 gene in E. histolytica isolates CONCLUSION: We developed a PCR method, which could detect simply and rapidly E. histolytica by amplifying a specific PCR fragment. |
format | Online Article Text |
id | pubmed-3481692 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Tehran University of Medical Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-34816922012-10-30 Evaluation of a Single PCR Assays on Cp5 Gene for Differentiation of Entamoeba histolytica and E. dispar Rostamighalehjaghi, S Jamali, R Rezaie, S Babaei, Z Hooshyar, H Rezaeian, M Iran J Public Health Original Article BACKGROUND: We examined a molecular method with a single-PCR for amplification of a part of CP5 gene enabling us to differentiate the pathogenic species, Entamoeba histolytica, from the non-pathogenic species, E. dispar. METHODS: We developed a single PCR method for this purpose. After investigation of GenBank, primer pairs were designed from highly conserved regions of cysteine proteinase (CP5) gene. The primers were utilized in PCR using isolated genomic DNA template of E. histolytica and the PCR products were then sequenced. The same primer and method for PCR was used for isolated genomic DNA template of E. dispar. RESULTS: A fragment of about 950 bp was isolated in PCR by using DNA from E. histolytica, however, no banding pattern was produced by using the same primers for E. dispar. We characterized CP5 gene at molecular level in E. histolytica isolates from 22 positive; including 20 non-dysentery samples isolated from both cities as well as two dysentery samples isolated only from Tabriz. Nucleotide sequence comparison in gene data banks (NCBI, NIH) revealed significant homology with CP5 gene in E. histolytica isolates CONCLUSION: We developed a PCR method, which could detect simply and rapidly E. histolytica by amplifying a specific PCR fragment. Tehran University of Medical Sciences 2010-12-31 /pmc/articles/PMC3481692/ /pubmed/23113039 Text en Copyright © Iranian Public Health Association & Tehran University of Medical Sciences http://creativecommons.org/licenses/by-nc/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution NonCommercial 3.0 License (CC BY-NC 3.0), which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly. |
spellingShingle | Original Article Rostamighalehjaghi, S Jamali, R Rezaie, S Babaei, Z Hooshyar, H Rezaeian, M Evaluation of a Single PCR Assays on Cp5 Gene for Differentiation of Entamoeba histolytica and E. dispar |
title | Evaluation of a Single PCR Assays on Cp5 Gene for Differentiation of Entamoeba histolytica and E. dispar |
title_full | Evaluation of a Single PCR Assays on Cp5 Gene for Differentiation of Entamoeba histolytica and E. dispar |
title_fullStr | Evaluation of a Single PCR Assays on Cp5 Gene for Differentiation of Entamoeba histolytica and E. dispar |
title_full_unstemmed | Evaluation of a Single PCR Assays on Cp5 Gene for Differentiation of Entamoeba histolytica and E. dispar |
title_short | Evaluation of a Single PCR Assays on Cp5 Gene for Differentiation of Entamoeba histolytica and E. dispar |
title_sort | evaluation of a single pcr assays on cp5 gene for differentiation of entamoeba histolytica and e. dispar |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3481692/ https://www.ncbi.nlm.nih.gov/pubmed/23113039 |
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