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Application of Hybridization Chain Reaction/CRISPR-Cas12a for the Detection of SARS-CoV-2 Infection

Globally, the emergence of the coronavirus disease (COVID-19) has had a significant impact on life. The need for ongoing SARS-CoV-2 screening employing inexpensive and quick diagnostic approaches is undeniable, given the ongoing pandemic and variations in vaccine administration in resource-constrain...

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Autores principales: Sagoe, Kate Obaayaa, Kyama, Mutinda Cleophas, Maina, Naomi, Kamita, Moses, Njokah, Muturi, Thiong’o, Kelvin, Kanoi, Bernard N., Wandera, Ernest Apondi, Ndegwa, Davies, Kinyua, Dickson Mwenda, Gitaka, Jesse
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10178590/
https://www.ncbi.nlm.nih.gov/pubmed/37175035
http://dx.doi.org/10.3390/diagnostics13091644
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author Sagoe, Kate Obaayaa
Kyama, Mutinda Cleophas
Maina, Naomi
Kamita, Moses
Njokah, Muturi
Thiong’o, Kelvin
Kanoi, Bernard N.
Wandera, Ernest Apondi
Ndegwa, Davies
Kinyua, Dickson Mwenda
Gitaka, Jesse
author_facet Sagoe, Kate Obaayaa
Kyama, Mutinda Cleophas
Maina, Naomi
Kamita, Moses
Njokah, Muturi
Thiong’o, Kelvin
Kanoi, Bernard N.
Wandera, Ernest Apondi
Ndegwa, Davies
Kinyua, Dickson Mwenda
Gitaka, Jesse
author_sort Sagoe, Kate Obaayaa
collection PubMed
description Globally, the emergence of the coronavirus disease (COVID-19) has had a significant impact on life. The need for ongoing SARS-CoV-2 screening employing inexpensive and quick diagnostic approaches is undeniable, given the ongoing pandemic and variations in vaccine administration in resource-constrained regions. This study presents results as proof of concept to use hybridization chain reaction (HCR) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a complex for detecting SARS-CoV-2. HCR hairpin probes were designed using the NUPACK web-based program and further used to amplify the SARS-CoV-2 N gene in archived nasopharyngeal samples. The results were visualized using agarose gels and CRISPR Cas12a-based lateral flow strips. The assay was evaluated using the gold standard, real-time polymerase chain reaction (RT-PCR), as recommended by the World Health Organization (WHO). The results show the comparative efficiency of HCR to RT-PCR. This study shows that HCR and CRISPR are viable alternatives for diagnosing SARS-CoV-2 in samples.
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spelling pubmed-101785902023-05-13 Application of Hybridization Chain Reaction/CRISPR-Cas12a for the Detection of SARS-CoV-2 Infection Sagoe, Kate Obaayaa Kyama, Mutinda Cleophas Maina, Naomi Kamita, Moses Njokah, Muturi Thiong’o, Kelvin Kanoi, Bernard N. Wandera, Ernest Apondi Ndegwa, Davies Kinyua, Dickson Mwenda Gitaka, Jesse Diagnostics (Basel) Article Globally, the emergence of the coronavirus disease (COVID-19) has had a significant impact on life. The need for ongoing SARS-CoV-2 screening employing inexpensive and quick diagnostic approaches is undeniable, given the ongoing pandemic and variations in vaccine administration in resource-constrained regions. This study presents results as proof of concept to use hybridization chain reaction (HCR) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a complex for detecting SARS-CoV-2. HCR hairpin probes were designed using the NUPACK web-based program and further used to amplify the SARS-CoV-2 N gene in archived nasopharyngeal samples. The results were visualized using agarose gels and CRISPR Cas12a-based lateral flow strips. The assay was evaluated using the gold standard, real-time polymerase chain reaction (RT-PCR), as recommended by the World Health Organization (WHO). The results show the comparative efficiency of HCR to RT-PCR. This study shows that HCR and CRISPR are viable alternatives for diagnosing SARS-CoV-2 in samples. MDPI 2023-05-07 /pmc/articles/PMC10178590/ /pubmed/37175035 http://dx.doi.org/10.3390/diagnostics13091644 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Sagoe, Kate Obaayaa
Kyama, Mutinda Cleophas
Maina, Naomi
Kamita, Moses
Njokah, Muturi
Thiong’o, Kelvin
Kanoi, Bernard N.
Wandera, Ernest Apondi
Ndegwa, Davies
Kinyua, Dickson Mwenda
Gitaka, Jesse
Application of Hybridization Chain Reaction/CRISPR-Cas12a for the Detection of SARS-CoV-2 Infection
title Application of Hybridization Chain Reaction/CRISPR-Cas12a for the Detection of SARS-CoV-2 Infection
title_full Application of Hybridization Chain Reaction/CRISPR-Cas12a for the Detection of SARS-CoV-2 Infection
title_fullStr Application of Hybridization Chain Reaction/CRISPR-Cas12a for the Detection of SARS-CoV-2 Infection
title_full_unstemmed Application of Hybridization Chain Reaction/CRISPR-Cas12a for the Detection of SARS-CoV-2 Infection
title_short Application of Hybridization Chain Reaction/CRISPR-Cas12a for the Detection of SARS-CoV-2 Infection
title_sort application of hybridization chain reaction/crispr-cas12a for the detection of sars-cov-2 infection
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10178590/
https://www.ncbi.nlm.nih.gov/pubmed/37175035
http://dx.doi.org/10.3390/diagnostics13091644
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