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Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction
This study aimed to detect the SARS-COV2 viral component directly from inoculated VTM without RNA extraction. Inoculated VTMs of already tested 50 positive and 50 negative samples were divided into three groups. Group I was treated with Proteinase K (PK) followed by 3-step-heat treatment at differen...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8595283/ https://www.ncbi.nlm.nih.gov/pubmed/34804989 http://dx.doi.org/10.3389/fcimb.2021.717068 |
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author | Shukla, Alka Gangwar, Mayank Sharma, Gaurav Prakash, Pradyot Nath, Gopal |
author_facet | Shukla, Alka Gangwar, Mayank Sharma, Gaurav Prakash, Pradyot Nath, Gopal |
author_sort | Shukla, Alka |
collection | PubMed |
description | This study aimed to detect the SARS-COV2 viral component directly from inoculated VTM without RNA extraction. Inoculated VTMs of already tested 50 positive and 50 negative samples were divided into three groups. Group I was treated with Proteinase K (PK) followed by 3-step-heat treatment at different temperatures (25°C, 60°C, and 98°C) and stored at 4°C. Group II was directly subjected to 3-step-heat treatment without PK exposure and stored at 4°C. And group III was set-up as standard group; it was processed using Qiagen’s column based QIAamp Nucleic Acid kit and the obtained nucleic acids were stored at 4°C. These stored samples were used as a template to execute real-time polymerase chain reaction, and results were noted. Group I demonstrated 96% and 88% sensitivity for N and ORF1ab genes respectively, whereas group II demonstrated 78% and 60% when compared to the results of standard group III. Overall group I showed better results than group II when compared to group III. Thus, in situations where gold-standard reagents are not available, PK exposure and heat treatment can be employed to carry out molecular detection of SARS-CoV2 viral component. |
format | Online Article Text |
id | pubmed-8595283 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-85952832021-11-18 Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction Shukla, Alka Gangwar, Mayank Sharma, Gaurav Prakash, Pradyot Nath, Gopal Front Cell Infect Microbiol Cellular and Infection Microbiology This study aimed to detect the SARS-COV2 viral component directly from inoculated VTM without RNA extraction. Inoculated VTMs of already tested 50 positive and 50 negative samples were divided into three groups. Group I was treated with Proteinase K (PK) followed by 3-step-heat treatment at different temperatures (25°C, 60°C, and 98°C) and stored at 4°C. Group II was directly subjected to 3-step-heat treatment without PK exposure and stored at 4°C. And group III was set-up as standard group; it was processed using Qiagen’s column based QIAamp Nucleic Acid kit and the obtained nucleic acids were stored at 4°C. These stored samples were used as a template to execute real-time polymerase chain reaction, and results were noted. Group I demonstrated 96% and 88% sensitivity for N and ORF1ab genes respectively, whereas group II demonstrated 78% and 60% when compared to the results of standard group III. Overall group I showed better results than group II when compared to group III. Thus, in situations where gold-standard reagents are not available, PK exposure and heat treatment can be employed to carry out molecular detection of SARS-CoV2 viral component. Frontiers Media S.A. 2021-11-03 /pmc/articles/PMC8595283/ /pubmed/34804989 http://dx.doi.org/10.3389/fcimb.2021.717068 Text en Copyright © 2021 Shukla, Gangwar, Sharma, Prakash and Nath https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Cellular and Infection Microbiology Shukla, Alka Gangwar, Mayank Sharma, Gaurav Prakash, Pradyot Nath, Gopal Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction |
title | Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction |
title_full | Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction |
title_fullStr | Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction |
title_full_unstemmed | Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction |
title_short | Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction |
title_sort | vitality of proteinase k in rrtpcr detection of sars-cov2 bypassing rna extraction |
topic | Cellular and Infection Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8595283/ https://www.ncbi.nlm.nih.gov/pubmed/34804989 http://dx.doi.org/10.3389/fcimb.2021.717068 |
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