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Isothermal PCR for Feasible Molecular Diagnosis of Primary Toxoplasmosis in Women Recently Experienced Spontaneous Abortion

AIM: The current study aimed to assess the practicability of a simple loop-mediated isothermal amplification (LAMP) about real-time quantitative PCR to diagnose primary toxoplasmosis among high-risk pregnant women. METHODS: Cloned Toxoplasma samples were used to calculate the analytical sensitivity...

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Autores principales: El Aal, Amany A. Abd, Nahnoush, Reham K., Elmallawany, Marwa A., El-Sherbiny, Walid S., 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/PMC6026412/
https://www.ncbi.nlm.nih.gov/pubmed/29983788
http://dx.doi.org/10.3889/oamjms.2018.227
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author El Aal, Amany A. Abd
Nahnoush, Reham K.
Elmallawany, Marwa A.
El-Sherbiny, Walid S.
Badr, Mohamed S.
Nasr, Ghada M.
author_facet El Aal, Amany A. Abd
Nahnoush, Reham K.
Elmallawany, Marwa A.
El-Sherbiny, Walid S.
Badr, Mohamed S.
Nasr, Ghada M.
author_sort El Aal, Amany A. Abd
collection PubMed
description AIM: The current study aimed to assess the practicability of a simple loop-mediated isothermal amplification (LAMP) about real-time quantitative PCR to diagnose primary toxoplasmosis among high-risk pregnant women. METHODS: Cloned Toxoplasma samples were used to calculate the analytical sensitivity while specificity was assessed using pooled DNA samples extracted from other parasitic stages. RESULTS: Both techniques showed 100% sensitivity and specificity and then applied to detect recent Toxoplasma infection in peripheral blood of 77 IgG negative women out of a total 139 women lately experienced spontaneous abortion. The 2 techniques obtained positive results in 8 samples confirming primary toxoplasmosis. CONCLUSION: Generally, LAMP assay is a simple, cost-effective molecular technique can be completed in less than half an hour to diagnose primary Toxoplasma infection. The technique can be applied in a minimally equipped laboratory by ordinary workers to screen the vulnerable groups. Further analysis using larger samples with the quantitative approach is recommended to confirm the sensitivity of this emergent molecular technique.
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spelling pubmed-60264122018-07-06 Isothermal PCR for Feasible Molecular Diagnosis of Primary Toxoplasmosis in Women Recently Experienced Spontaneous Abortion El Aal, Amany A. Abd Nahnoush, Reham K. Elmallawany, Marwa A. El-Sherbiny, Walid S. Badr, Mohamed S. Nasr, Ghada M. Open Access Maced J Med Sci Basic Science AIM: The current study aimed to assess the practicability of a simple loop-mediated isothermal amplification (LAMP) about real-time quantitative PCR to diagnose primary toxoplasmosis among high-risk pregnant women. METHODS: Cloned Toxoplasma samples were used to calculate the analytical sensitivity while specificity was assessed using pooled DNA samples extracted from other parasitic stages. RESULTS: Both techniques showed 100% sensitivity and specificity and then applied to detect recent Toxoplasma infection in peripheral blood of 77 IgG negative women out of a total 139 women lately experienced spontaneous abortion. The 2 techniques obtained positive results in 8 samples confirming primary toxoplasmosis. CONCLUSION: Generally, LAMP assay is a simple, cost-effective molecular technique can be completed in less than half an hour to diagnose primary Toxoplasma infection. The technique can be applied in a minimally equipped laboratory by ordinary workers to screen the vulnerable groups. Further analysis using larger samples with the quantitative approach is recommended to confirm the sensitivity of this emergent molecular technique. Republic of Macedonia 2018-06-10 /pmc/articles/PMC6026412/ /pubmed/29983788 http://dx.doi.org/10.3889/oamjms.2018.227 Text en Copyright: © 2018 Amany A. Abd El Aal, Reham K. Nahnoush, Marwa A. Elmallawany, Walid S. El-Sherbiny, 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
El Aal, Amany A. Abd
Nahnoush, Reham K.
Elmallawany, Marwa A.
El-Sherbiny, Walid S.
Badr, Mohamed S.
Nasr, Ghada M.
Isothermal PCR for Feasible Molecular Diagnosis of Primary Toxoplasmosis in Women Recently Experienced Spontaneous Abortion
title Isothermal PCR for Feasible Molecular Diagnosis of Primary Toxoplasmosis in Women Recently Experienced Spontaneous Abortion
title_full Isothermal PCR for Feasible Molecular Diagnosis of Primary Toxoplasmosis in Women Recently Experienced Spontaneous Abortion
title_fullStr Isothermal PCR for Feasible Molecular Diagnosis of Primary Toxoplasmosis in Women Recently Experienced Spontaneous Abortion
title_full_unstemmed Isothermal PCR for Feasible Molecular Diagnosis of Primary Toxoplasmosis in Women Recently Experienced Spontaneous Abortion
title_short Isothermal PCR for Feasible Molecular Diagnosis of Primary Toxoplasmosis in Women Recently Experienced Spontaneous Abortion
title_sort isothermal pcr for feasible molecular diagnosis of primary toxoplasmosis in women recently experienced spontaneous abortion
topic Basic Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6026412/
https://www.ncbi.nlm.nih.gov/pubmed/29983788
http://dx.doi.org/10.3889/oamjms.2018.227
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