4.6 Article

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

Journal

DIAGNOSTICS
Volume 13, Issue 9, Pages -

Publisher

MDPI
DOI: 10.3390/diagnostics13091644

Keywords

hybridization chain reaction (HCR); severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2); clustered regularly interspaced short palindromic repeats (CRISPR); COVID-19; point-of-care (POC); diagnostics

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This study presents a proof of concept for using hybridization chain reaction (HCR) and clustered regularly interspaces short palindromic repeats (CRISPR)/Cas12a complex to detect SARS-CoV-2. The results show the comparative efficiency of HCR to RT-PCR, indicating that HCR and CRISPR are viable alternatives for diagnosing SARS-CoV-2 in samples.
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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