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16S rRNA Monitoring Point-of-Care Magnetic Focus Lateral Flow Sensor
[Image: see text] The detection and profiling of pathogenic bacteria is critical for human health, environmental, and food safety monitoring. Herein, we propose a highly sensitive colorimetric strategy for naked eye screening of 16S ribosomal RNA (16S rRNA) from pathogenic agents relevant to infecti...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8153928/ https://www.ncbi.nlm.nih.gov/pubmed/34056264 http://dx.doi.org/10.1021/acsomega.1c01307 |
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author | Ren, Wen Ahmad, Saeed Irudayaraj, Joseph |
author_facet | Ren, Wen Ahmad, Saeed Irudayaraj, Joseph |
author_sort | Ren, Wen |
collection | PubMed |
description | [Image: see text] The detection and profiling of pathogenic bacteria is critical for human health, environmental, and food safety monitoring. Herein, we propose a highly sensitive colorimetric strategy for naked eye screening of 16S ribosomal RNA (16S rRNA) from pathogenic agents relevant to infections, human health, and food safety monitoring with a magnetic focus lateral flow sensor (mLFS) platform. The method developed was demonstrated in model 16S rRNA sequences of the pathogen Escherichia coli O157:H7 to detect as low as 1 fM of targets, exhibiting a sensitivity improved by ∼5 × 10(5) times compared to the conventional GNP-based colorimetric lateral flow assay used for oligonucleotide testing. Based on the grayscale values, semi-quantitation of up to 1 pM of target sequences was possible in ∼45 min. The methodology could detect the target 16S rRNA from as low as 32 pg/mL of total RNA extracted from pathogens. Specificity was demonstrated with total RNA extracted from E. coli K-12 MG1655, Bacillus subtilis (B. subtilis), and Pseudomonas aeruginosa (P. aeruginosa). No signal was observed from as high as 320 pg/mL of total RNA from the nontarget bacteria. The recognition of target 16S rRNA from 32 pg/mL of total RNA in complex matrices was also demonstrated. The proposed mLFS method was then extended to monitoring B. subtilis and P. aeruginosa. Our approach highlights the possibility of extending this concept to screen specific nucleic acid sequences for the monitoring of infectious pathogens or microbiome implicated in a range of diseases including cancer. |
format | Online Article Text |
id | pubmed-8153928 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-81539282021-05-27 16S rRNA Monitoring Point-of-Care Magnetic Focus Lateral Flow Sensor Ren, Wen Ahmad, Saeed Irudayaraj, Joseph ACS Omega [Image: see text] The detection and profiling of pathogenic bacteria is critical for human health, environmental, and food safety monitoring. Herein, we propose a highly sensitive colorimetric strategy for naked eye screening of 16S ribosomal RNA (16S rRNA) from pathogenic agents relevant to infections, human health, and food safety monitoring with a magnetic focus lateral flow sensor (mLFS) platform. The method developed was demonstrated in model 16S rRNA sequences of the pathogen Escherichia coli O157:H7 to detect as low as 1 fM of targets, exhibiting a sensitivity improved by ∼5 × 10(5) times compared to the conventional GNP-based colorimetric lateral flow assay used for oligonucleotide testing. Based on the grayscale values, semi-quantitation of up to 1 pM of target sequences was possible in ∼45 min. The methodology could detect the target 16S rRNA from as low as 32 pg/mL of total RNA extracted from pathogens. Specificity was demonstrated with total RNA extracted from E. coli K-12 MG1655, Bacillus subtilis (B. subtilis), and Pseudomonas aeruginosa (P. aeruginosa). No signal was observed from as high as 320 pg/mL of total RNA from the nontarget bacteria. The recognition of target 16S rRNA from 32 pg/mL of total RNA in complex matrices was also demonstrated. The proposed mLFS method was then extended to monitoring B. subtilis and P. aeruginosa. Our approach highlights the possibility of extending this concept to screen specific nucleic acid sequences for the monitoring of infectious pathogens or microbiome implicated in a range of diseases including cancer. American Chemical Society 2021-04-14 /pmc/articles/PMC8153928/ /pubmed/34056264 http://dx.doi.org/10.1021/acsomega.1c01307 Text en © 2021 The Authors. Published by American Chemical Society Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Ren, Wen Ahmad, Saeed Irudayaraj, Joseph 16S rRNA Monitoring Point-of-Care Magnetic Focus Lateral Flow Sensor |
title | 16S rRNA Monitoring Point-of-Care Magnetic Focus Lateral
Flow Sensor |
title_full | 16S rRNA Monitoring Point-of-Care Magnetic Focus Lateral
Flow Sensor |
title_fullStr | 16S rRNA Monitoring Point-of-Care Magnetic Focus Lateral
Flow Sensor |
title_full_unstemmed | 16S rRNA Monitoring Point-of-Care Magnetic Focus Lateral
Flow Sensor |
title_short | 16S rRNA Monitoring Point-of-Care Magnetic Focus Lateral
Flow Sensor |
title_sort | 16s rrna monitoring point-of-care magnetic focus lateral
flow sensor |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8153928/ https://www.ncbi.nlm.nih.gov/pubmed/34056264 http://dx.doi.org/10.1021/acsomega.1c01307 |
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