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Performance of 2 Polymerization Protocols Targeting Cloned Toxoplasma Parasites

BACKGROUND: Toxoplasma gondii is a common parasitic infection of humans. Infection is usually mild. Serious complications can occur in pregnant and immunocompromised patients. AIM: The present study aims to investigate the performance of 2 different PCR protocols; real-time quantitative molecular as...

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Autores principales: Hanafy, Nihal A., Badr, Mohamed S., Nasr, Ghada M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Republic of Macedonia 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6182548/
https://www.ncbi.nlm.nih.gov/pubmed/30337968
http://dx.doi.org/10.3889/oamjms.2018.400
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author Hanafy, Nihal A.
Badr, Mohamed S.
Nasr, Ghada M.
author_facet Hanafy, Nihal A.
Badr, Mohamed S.
Nasr, Ghada M.
author_sort Hanafy, Nihal A.
collection PubMed
description BACKGROUND: Toxoplasma gondii is a common parasitic infection of humans. Infection is usually mild. Serious complications can occur in pregnant and immunocompromised patients. AIM: The present study aims to investigate the performance of 2 different PCR protocols; real-time quantitative molecular assays (qPCR) and conventional molecular assays (cPCR), using 2 different sets of primers and by using cloned purified Toxoplasma genomic substances to be evaluated as reference samples. METHODS: The target DNA was provided in 8 different quantities. RESULTS: Amplification failure was reported only with the cPCR in samples of low concentrations using both primer sets. Quantitative PCR detected the 8 different dilutions of the purified Toxoplasma gondii using the 2 sets of primers while cPCR was sensitive to detect only 6 different dilutions. CONCLUSION: Generally real-time quantitative molecular assays, is easy to use method compared to conventional PCR assay and produces more reliable results within only one hour time but still the possible application of qPCRs in routine diagnosis necessitates analysis of a large number of clinical samples in further studies to make the proper choice.
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spelling pubmed-61825482018-10-18 Performance of 2 Polymerization Protocols Targeting Cloned Toxoplasma Parasites Hanafy, Nihal A. Badr, Mohamed S. Nasr, Ghada M. Open Access Maced J Med Sci Basic Science BACKGROUND: Toxoplasma gondii is a common parasitic infection of humans. Infection is usually mild. Serious complications can occur in pregnant and immunocompromised patients. AIM: The present study aims to investigate the performance of 2 different PCR protocols; real-time quantitative molecular assays (qPCR) and conventional molecular assays (cPCR), using 2 different sets of primers and by using cloned purified Toxoplasma genomic substances to be evaluated as reference samples. METHODS: The target DNA was provided in 8 different quantities. RESULTS: Amplification failure was reported only with the cPCR in samples of low concentrations using both primer sets. Quantitative PCR detected the 8 different dilutions of the purified Toxoplasma gondii using the 2 sets of primers while cPCR was sensitive to detect only 6 different dilutions. CONCLUSION: Generally real-time quantitative molecular assays, is easy to use method compared to conventional PCR assay and produces more reliable results within only one hour time but still the possible application of qPCRs in routine diagnosis necessitates analysis of a large number of clinical samples in further studies to make the proper choice. Republic of Macedonia 2018-09-22 /pmc/articles/PMC6182548/ /pubmed/30337968 http://dx.doi.org/10.3889/oamjms.2018.400 Text en Copyright: © 2018 Nihal A. Hanafy, Mohamed S. Badr, Ghada M. Nasr. http://creativecommons.org/licenses/CC BY-NC/4.0 This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (CC BY-NC 4.0).
spellingShingle Basic Science
Hanafy, Nihal A.
Badr, Mohamed S.
Nasr, Ghada M.
Performance of 2 Polymerization Protocols Targeting Cloned Toxoplasma Parasites
title Performance of 2 Polymerization Protocols Targeting Cloned Toxoplasma Parasites
title_full Performance of 2 Polymerization Protocols Targeting Cloned Toxoplasma Parasites
title_fullStr Performance of 2 Polymerization Protocols Targeting Cloned Toxoplasma Parasites
title_full_unstemmed Performance of 2 Polymerization Protocols Targeting Cloned Toxoplasma Parasites
title_short Performance of 2 Polymerization Protocols Targeting Cloned Toxoplasma Parasites
title_sort performance of 2 polymerization protocols targeting cloned toxoplasma parasites
topic Basic Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6182548/
https://www.ncbi.nlm.nih.gov/pubmed/30337968
http://dx.doi.org/10.3889/oamjms.2018.400
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