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Development of RNA-Based Assay for Rapid Detection of SARS-CoV-2 in Clinical Samples

INTRODUCTION: The ongoing spread of pandemic coronavirus disease-19 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is of growing concern. Rapid diagnosis and management of SARS-CoV-2 are crucial for controlling the outbreak in the community. Here, we report the dev...

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Autores principales: Kumar, Vinod, Mishra, Suman, Sharma, Rajni, Agarwal, Jyotsna, Ghoshal, Ujjala, Khanna, Tripti, Sharma, Lokendra K., Verma, Santosh Kumar, Mishra, Prabhakar, Tiwari, Swasti
Formato: Online Artículo Texto
Lenguaje:English
Publicado: S. Karger AG 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9393769/
https://www.ncbi.nlm.nih.gov/pubmed/35193136
http://dx.doi.org/10.1159/000522337
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author Kumar, Vinod
Mishra, Suman
Sharma, Rajni
Agarwal, Jyotsna
Ghoshal, Ujjala
Khanna, Tripti
Sharma, Lokendra K.
Verma, Santosh Kumar
Mishra, Prabhakar
Tiwari, Swasti
author_facet Kumar, Vinod
Mishra, Suman
Sharma, Rajni
Agarwal, Jyotsna
Ghoshal, Ujjala
Khanna, Tripti
Sharma, Lokendra K.
Verma, Santosh Kumar
Mishra, Prabhakar
Tiwari, Swasti
author_sort Kumar, Vinod
collection PubMed
description INTRODUCTION: The ongoing spread of pandemic coronavirus disease-19 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is of growing concern. Rapid diagnosis and management of SARS-CoV-2 are crucial for controlling the outbreak in the community. Here, we report the development of a first rapid-colorimetric assay capable of detecting SARS-CoV-2 in the human nasopharyngeal RNA sample in less than 30 min. METHOD: We utilized a nanomaterial-based optical sensing platform to detect RNA-dependent RNA polymerase gene of SARS-CoV-2, where the formation of oligo probe-target hybrid led to salt-induced aggregation and change in gold-colloid color from pink to blue visibility range. Accordingly, we found a change in colloid color from pink to blue in assay containing nasopharyngeal RNA sample from the subject with clinically diagnosed COVID-19. The colloid retained pink color when the test includes samples from COVID-19 negative subjects or human papillomavirus-infected women. RESULTS: The results were validated using nasopharyngeal RNA samples from positive COVID-19 subjects (n = 136). Using real-time polymerase chain reaction as gold standard, the assay was found to have 85.29% sensitivity and 94.12% specificity. The optimized method has detection limit as little as 0.5 ng of SARS-CoV-2 RNA. CONCLUSION: We found that the developed assay rapidly detects SARS-CoV-2 RNA in clinical samples in a cost-effective manner and would be useful in pandemic management by facilitating mass screening.
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spelling pubmed-93937692022-08-22 Development of RNA-Based Assay for Rapid Detection of SARS-CoV-2 in Clinical Samples Kumar, Vinod Mishra, Suman Sharma, Rajni Agarwal, Jyotsna Ghoshal, Ujjala Khanna, Tripti Sharma, Lokendra K. Verma, Santosh Kumar Mishra, Prabhakar Tiwari, Swasti Intervirology Research Article INTRODUCTION: The ongoing spread of pandemic coronavirus disease-19 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is of growing concern. Rapid diagnosis and management of SARS-CoV-2 are crucial for controlling the outbreak in the community. Here, we report the development of a first rapid-colorimetric assay capable of detecting SARS-CoV-2 in the human nasopharyngeal RNA sample in less than 30 min. METHOD: We utilized a nanomaterial-based optical sensing platform to detect RNA-dependent RNA polymerase gene of SARS-CoV-2, where the formation of oligo probe-target hybrid led to salt-induced aggregation and change in gold-colloid color from pink to blue visibility range. Accordingly, we found a change in colloid color from pink to blue in assay containing nasopharyngeal RNA sample from the subject with clinically diagnosed COVID-19. The colloid retained pink color when the test includes samples from COVID-19 negative subjects or human papillomavirus-infected women. RESULTS: The results were validated using nasopharyngeal RNA samples from positive COVID-19 subjects (n = 136). Using real-time polymerase chain reaction as gold standard, the assay was found to have 85.29% sensitivity and 94.12% specificity. The optimized method has detection limit as little as 0.5 ng of SARS-CoV-2 RNA. CONCLUSION: We found that the developed assay rapidly detects SARS-CoV-2 RNA in clinical samples in a cost-effective manner and would be useful in pandemic management by facilitating mass screening. S. Karger AG 2022-02-22 /pmc/articles/PMC9393769/ /pubmed/35193136 http://dx.doi.org/10.1159/000522337 Text en Copyright © 2022 by The Author(s). Published by S. Karger AG, Basel https://creativecommons.org/licenses/by-nc/4.0/This article is licensed under the Creative Commons Attribution-NonCommercial 4.0 International License (CC BY-NC). Usage and distribution for commercial purposes requires written permission.
spellingShingle Research Article
Kumar, Vinod
Mishra, Suman
Sharma, Rajni
Agarwal, Jyotsna
Ghoshal, Ujjala
Khanna, Tripti
Sharma, Lokendra K.
Verma, Santosh Kumar
Mishra, Prabhakar
Tiwari, Swasti
Development of RNA-Based Assay for Rapid Detection of SARS-CoV-2 in Clinical Samples
title Development of RNA-Based Assay for Rapid Detection of SARS-CoV-2 in Clinical Samples
title_full Development of RNA-Based Assay for Rapid Detection of SARS-CoV-2 in Clinical Samples
title_fullStr Development of RNA-Based Assay for Rapid Detection of SARS-CoV-2 in Clinical Samples
title_full_unstemmed Development of RNA-Based Assay for Rapid Detection of SARS-CoV-2 in Clinical Samples
title_short Development of RNA-Based Assay for Rapid Detection of SARS-CoV-2 in Clinical Samples
title_sort development of rna-based assay for rapid detection of sars-cov-2 in clinical samples
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9393769/
https://www.ncbi.nlm.nih.gov/pubmed/35193136
http://dx.doi.org/10.1159/000522337
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