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Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii
BACKGROUND: Entamoeba moshkovskii and E. dispar are impossible to differentiate microscopically from the pathogenic species E. histolytica. Multiplex polymerase chain reaction (Multiplex PCR) is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Tehran University of Medical Sciences
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4345085/ https://www.ncbi.nlm.nih.gov/pubmed/25759727 |
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author | ZEBARDAST, Nozhat HAGHIGHI, Ali YEGANEH, Farshid SEYYED TABAEI, Seyyed Javad GHARAVI, Mohammad Javad FALLAHI, Shirzad LASJERDI, Zohreh SALEHI, Nima TAGHIPOUR, Niloofar KOHANSAL, Cobra NADERI, Farideh |
author_facet | ZEBARDAST, Nozhat HAGHIGHI, Ali YEGANEH, Farshid SEYYED TABAEI, Seyyed Javad GHARAVI, Mohammad Javad FALLAHI, Shirzad LASJERDI, Zohreh SALEHI, Nima TAGHIPOUR, Niloofar KOHANSAL, Cobra NADERI, Farideh |
author_sort | ZEBARDAST, Nozhat |
collection | PubMed |
description | BACKGROUND: Entamoeba moshkovskii and E. dispar are impossible to differentiate microscopically from the pathogenic species E. histolytica. Multiplex polymerase chain reaction (Multiplex PCR) is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment. METHODS: For detection and differentiation of the three-microscopy indistinguishable Entamoeba species in human, multiplex PCR assay using different DNA extraction methods was studied. A conserved forward primer was derived from the middle of the small-subunit rRNA gene, and reverse primers were designed from signature sequences specific to each of these three Entamoeba species. RESULTS: A 166-bp PCR product with E. histolytica DNA, a 580-bp product with E. moshkovskii DNA and a 752-bp product with E. dispar DNA were generated in a single-round and multiplex PCR reaction. CONCLUSION: We recommend this PCR assay as an accurate, rapid, and effective diagnostic method for the detection and discrimination of these three Entamoeba species in both routine diagnosis of amoebiasis and epidemiological surveys. |
format | Online Article Text |
id | pubmed-4345085 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Tehran University of Medical Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-43450852015-03-10 Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii ZEBARDAST, Nozhat HAGHIGHI, Ali YEGANEH, Farshid SEYYED TABAEI, Seyyed Javad GHARAVI, Mohammad Javad FALLAHI, Shirzad LASJERDI, Zohreh SALEHI, Nima TAGHIPOUR, Niloofar KOHANSAL, Cobra NADERI, Farideh Iran J Parasitol Medical Sciences BACKGROUND: Entamoeba moshkovskii and E. dispar are impossible to differentiate microscopically from the pathogenic species E. histolytica. Multiplex polymerase chain reaction (Multiplex PCR) is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment. METHODS: For detection and differentiation of the three-microscopy indistinguishable Entamoeba species in human, multiplex PCR assay using different DNA extraction methods was studied. A conserved forward primer was derived from the middle of the small-subunit rRNA gene, and reverse primers were designed from signature sequences specific to each of these three Entamoeba species. RESULTS: A 166-bp PCR product with E. histolytica DNA, a 580-bp product with E. moshkovskii DNA and a 752-bp product with E. dispar DNA were generated in a single-round and multiplex PCR reaction. CONCLUSION: We recommend this PCR assay as an accurate, rapid, and effective diagnostic method for the detection and discrimination of these three Entamoeba species in both routine diagnosis of amoebiasis and epidemiological surveys. Tehran University of Medical Sciences 2014 /pmc/articles/PMC4345085/ /pubmed/25759727 Text en Copyright: © Iranian Journal of Parasitology & Tehran University of Medical Sciences This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License 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 | Medical Sciences ZEBARDAST, Nozhat HAGHIGHI, Ali YEGANEH, Farshid SEYYED TABAEI, Seyyed Javad GHARAVI, Mohammad Javad FALLAHI, Shirzad LASJERDI, Zohreh SALEHI, Nima TAGHIPOUR, Niloofar KOHANSAL, Cobra NADERI, Farideh Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii |
title | Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii |
title_full | Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii |
title_fullStr | Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii |
title_full_unstemmed | Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii |
title_short | Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii |
title_sort | application of multiplex pcr for detection and differentiation of entamoeba histolytica, entamoeba dispar and entamoeba moshkovskii |
topic | Medical Sciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4345085/ https://www.ncbi.nlm.nih.gov/pubmed/25759727 |
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