Visual Detection of SARS-CoV-2 RNA by Conventional PCR-Induced Generation of DNAzyme Sensor
The gold standard for the diagnosis of SARS-CoV-2, the causative agent of COVID-19, is real-time polymerase chain reaction (PCR), which is labor-intensive, expensive, and not widely available in resource-poor settings. Therefore, it is imperative to develop novel, accurate, affordable, and easily ac...
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doaj-f950b389befc4c7388f75c31eb8ddb732020-12-23T07:44:06ZengFrontiers Media S.A.Frontiers in Molecular Biosciences2296-889X2020-12-01710.3389/fmolb.2020.586254586254Visual Detection of SARS-CoV-2 RNA by Conventional PCR-Induced Generation of DNAzyme SensorAnbalagan Anantharaj0Soon Jyoti Das1Patil Sharanabasava2Rakesh Lodha3Sushil K. Kabra4Tarun Kumar Sharma5Guruprasad R. Medigeshi6National Capital Region - Biotech Science Cluster, Translational Health Science and Technology Institute (THSTI), Faridabad, IndiaNational Capital Region - Biotech Science Cluster, Translational Health Science and Technology Institute (THSTI), Faridabad, IndiaNational Capital Region - Biotech Science Cluster, Translational Health Science and Technology Institute (THSTI), Faridabad, IndiaDepartment of Pediatrics, All India Institute of Medical Sciences, New Delhi, IndiaDepartment of Pediatrics, All India Institute of Medical Sciences, New Delhi, IndiaNational Capital Region - Biotech Science Cluster, Translational Health Science and Technology Institute (THSTI), Faridabad, IndiaNational Capital Region - Biotech Science Cluster, Translational Health Science and Technology Institute (THSTI), Faridabad, IndiaThe gold standard for the diagnosis of SARS-CoV-2, the causative agent of COVID-19, is real-time polymerase chain reaction (PCR), which is labor-intensive, expensive, and not widely available in resource-poor settings. Therefore, it is imperative to develop novel, accurate, affordable, and easily accessible assays/sensors to diagnose and isolate COVID-19 cases. To address this unmet need, we utilized the catalytic potential of peroxidase-like DNAzyme and developed a simple visual detection assay for SARS-CoV-2 RNA using a conventional thermal cycler by the PCR-induced generation of DNAzyme sensor. The performance of RT-PCR DNAzyme-based sensor was comparable to that of real-time PCR. The pilot scale validation of RT-PCR DNAzyme-based sensor has shown ~100% sensitivity and specificity in clinical specimens (nasopharyngeal swab, n = 34), with a good correlation (Spearman r = 0.799) with the Ct-value of fluorescence probe-based real-time PCR. These findings clearly indicate the potential of this inexpensive, sensitive, and specific molecular diagnostic test to extend our testing capabilities for the detection of SARS-CoV-2 to curtail COVID-19 transmission.https://www.frontiersin.org/articles/10.3389/fmolb.2020.586254/fullcolorimetric assayCOVID-19SARS-CoV-2DNAzymesensorreal-time-PCR |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Anbalagan Anantharaj Soon Jyoti Das Patil Sharanabasava Rakesh Lodha Sushil K. Kabra Tarun Kumar Sharma Guruprasad R. Medigeshi |
spellingShingle |
Anbalagan Anantharaj Soon Jyoti Das Patil Sharanabasava Rakesh Lodha Sushil K. Kabra Tarun Kumar Sharma Guruprasad R. Medigeshi Visual Detection of SARS-CoV-2 RNA by Conventional PCR-Induced Generation of DNAzyme Sensor Frontiers in Molecular Biosciences colorimetric assay COVID-19 SARS-CoV-2 DNAzyme sensor real-time-PCR |
author_facet |
Anbalagan Anantharaj Soon Jyoti Das Patil Sharanabasava Rakesh Lodha Sushil K. Kabra Tarun Kumar Sharma Guruprasad R. Medigeshi |
author_sort |
Anbalagan Anantharaj |
title |
Visual Detection of SARS-CoV-2 RNA by Conventional PCR-Induced Generation of DNAzyme Sensor |
title_short |
Visual Detection of SARS-CoV-2 RNA by Conventional PCR-Induced Generation of DNAzyme Sensor |
title_full |
Visual Detection of SARS-CoV-2 RNA by Conventional PCR-Induced Generation of DNAzyme Sensor |
title_fullStr |
Visual Detection of SARS-CoV-2 RNA by Conventional PCR-Induced Generation of DNAzyme Sensor |
title_full_unstemmed |
Visual Detection of SARS-CoV-2 RNA by Conventional PCR-Induced Generation of DNAzyme Sensor |
title_sort |
visual detection of sars-cov-2 rna by conventional pcr-induced generation of dnazyme sensor |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Molecular Biosciences |
issn |
2296-889X |
publishDate |
2020-12-01 |
description |
The gold standard for the diagnosis of SARS-CoV-2, the causative agent of COVID-19, is real-time polymerase chain reaction (PCR), which is labor-intensive, expensive, and not widely available in resource-poor settings. Therefore, it is imperative to develop novel, accurate, affordable, and easily accessible assays/sensors to diagnose and isolate COVID-19 cases. To address this unmet need, we utilized the catalytic potential of peroxidase-like DNAzyme and developed a simple visual detection assay for SARS-CoV-2 RNA using a conventional thermal cycler by the PCR-induced generation of DNAzyme sensor. The performance of RT-PCR DNAzyme-based sensor was comparable to that of real-time PCR. The pilot scale validation of RT-PCR DNAzyme-based sensor has shown ~100% sensitivity and specificity in clinical specimens (nasopharyngeal swab, n = 34), with a good correlation (Spearman r = 0.799) with the Ct-value of fluorescence probe-based real-time PCR. These findings clearly indicate the potential of this inexpensive, sensitive, and specific molecular diagnostic test to extend our testing capabilities for the detection of SARS-CoV-2 to curtail COVID-19 transmission. |
topic |
colorimetric assay COVID-19 SARS-CoV-2 DNAzyme sensor real-time-PCR |
url |
https://www.frontiersin.org/articles/10.3389/fmolb.2020.586254/full |
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