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SARS-CoV-2 direct real-time polymerase chain reaction testing in laboratories with shortage challenges

Aim: In our study, we propose the use of direct real-time polymerase chain reaction (RT-PCR), this test does not require extraction or a preheating step, which saves a lot of cost, labor, processing time and provides a solution for supply shortage. Materials & methods: We assayed 185 nasopharyng...

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Autores principales: Alaifan, Taghreed, Altamimi, Asmaa, Obeid, Dalia, Alshehri, Turki, Almatrrouk, Shaihana, Albarrag, Ahmed
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Future Medicine Ltd 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7856914/
http://dx.doi.org/10.2217/fvl-2020-0187
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author Alaifan, Taghreed
Altamimi, Asmaa
Obeid, Dalia
Alshehri, Turki
Almatrrouk, Shaihana
Albarrag, Ahmed
author_facet Alaifan, Taghreed
Altamimi, Asmaa
Obeid, Dalia
Alshehri, Turki
Almatrrouk, Shaihana
Albarrag, Ahmed
author_sort Alaifan, Taghreed
collection PubMed
description Aim: In our study, we propose the use of direct real-time polymerase chain reaction (RT-PCR), this test does not require extraction or a preheating step, which saves a lot of cost, labor, processing time and provides a solution for supply shortage. Materials & methods: We assayed 185 nasopharyngeal samples stored in viral transport media. The indirect method was done with RNA extraction step, and the direct RT-PCR was done without an extraction step, both assays were evaluated on a commercially validated severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) kit targeting E gene. Results: Our assay showed a sensitivity of 79%, a specificity of 100% and the agreement between methods was 72%. Conclusion: Overall, this simple direct RT-PCR approach can be utilized as a qualitative diagnostic tool for emergency SARS-CoV-2 surveillance in countries with limited resources and may help laboratories to continue testing and at greater frequency despite supply shortages, although with delay in cycle threshold value in comparison with indirect RT-PCR.
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spelling pubmed-78569142021-02-03 SARS-CoV-2 direct real-time polymerase chain reaction testing in laboratories with shortage challenges Alaifan, Taghreed Altamimi, Asmaa Obeid, Dalia Alshehri, Turki Almatrrouk, Shaihana Albarrag, Ahmed Future Virol Research Article Aim: In our study, we propose the use of direct real-time polymerase chain reaction (RT-PCR), this test does not require extraction or a preheating step, which saves a lot of cost, labor, processing time and provides a solution for supply shortage. Materials & methods: We assayed 185 nasopharyngeal samples stored in viral transport media. The indirect method was done with RNA extraction step, and the direct RT-PCR was done without an extraction step, both assays were evaluated on a commercially validated severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) kit targeting E gene. Results: Our assay showed a sensitivity of 79%, a specificity of 100% and the agreement between methods was 72%. Conclusion: Overall, this simple direct RT-PCR approach can be utilized as a qualitative diagnostic tool for emergency SARS-CoV-2 surveillance in countries with limited resources and may help laboratories to continue testing and at greater frequency despite supply shortages, although with delay in cycle threshold value in comparison with indirect RT-PCR. Future Medicine Ltd 2021-02-03 2021-01 /pmc/articles/PMC7856914/ http://dx.doi.org/10.2217/fvl-2020-0187 Text en © 2021 Future Medicine Ltd This work is licensed under the Creative Commons Attribution 4.0 License (http://creativecommons.org/licenses/by/4.0/)
spellingShingle Research Article
Alaifan, Taghreed
Altamimi, Asmaa
Obeid, Dalia
Alshehri, Turki
Almatrrouk, Shaihana
Albarrag, Ahmed
SARS-CoV-2 direct real-time polymerase chain reaction testing in laboratories with shortage challenges
title SARS-CoV-2 direct real-time polymerase chain reaction testing in laboratories with shortage challenges
title_full SARS-CoV-2 direct real-time polymerase chain reaction testing in laboratories with shortage challenges
title_fullStr SARS-CoV-2 direct real-time polymerase chain reaction testing in laboratories with shortage challenges
title_full_unstemmed SARS-CoV-2 direct real-time polymerase chain reaction testing in laboratories with shortage challenges
title_short SARS-CoV-2 direct real-time polymerase chain reaction testing in laboratories with shortage challenges
title_sort sars-cov-2 direct real-time polymerase chain reaction testing in laboratories with shortage challenges
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7856914/
http://dx.doi.org/10.2217/fvl-2020-0187
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