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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Republic of Macedonia
2018
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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. |
format | Online Article Text |
id | pubmed-6026412 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Republic of Macedonia |
record_format | MEDLINE/PubMed |
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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