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Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles
Direct detection of genetic biomarkers in body fluid lysate without target amplification will revolutionize nucleic acid-based diagnostics. However, the low concentration of target sequences makes this goal challenging. We report a method for direct detection of pathogen RNA in blood lysate using a...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5840326/ https://www.ncbi.nlm.nih.gov/pubmed/29511216 http://dx.doi.org/10.1038/s41598-018-21615-3 |
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author | Ngo, Hoan T. Freedman, Elizabeth Odion, Ren Abelard Strobbia, Pietro De Silva Indrasekara, Agampodi Swarnapali Vohra, Priya Taylor, Steve M. Vo-Dinh, Tuan |
author_facet | Ngo, Hoan T. Freedman, Elizabeth Odion, Ren Abelard Strobbia, Pietro De Silva Indrasekara, Agampodi Swarnapali Vohra, Priya Taylor, Steve M. Vo-Dinh, Tuan |
author_sort | Ngo, Hoan T. |
collection | PubMed |
description | Direct detection of genetic biomarkers in body fluid lysate without target amplification will revolutionize nucleic acid-based diagnostics. However, the low concentration of target sequences makes this goal challenging. We report a method for direct detection of pathogen RNA in blood lysate using a bioassay using surface-enhanced Raman spectroscopy (SERS)-based detection integrated in a “lab-in-a-stick” portable device. Two levels of signal enhancement were employed to achieve the sensitivity required for direct detection. Each target sequence was tagged with an ultrabright SERS-encoded nanorattle with ultrahigh SERS signals, and these tagged target sequences were concentrated into a focused spot for detection using hybridization sandwiches with magnetic microbeads. Furthermore, the washing process was automated by integration into a “lab-in-a-stick” portable device. We could directly detect synthetic target with a limit of detection of 200 fM. More importantly, we detected plasmodium falciparum malaria parasite RNA directly in infected red blood cells lysate. To our knowledge, this is the first report of SERS-based direct detection of pathogen nucleic acid in blood lysate without nucleic acid extraction or target amplification. The results show the potential of our integrated bioassay for field use and point-of-care diagnostics. |
format | Online Article Text |
id | pubmed-5840326 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-58403262018-03-13 Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles Ngo, Hoan T. Freedman, Elizabeth Odion, Ren Abelard Strobbia, Pietro De Silva Indrasekara, Agampodi Swarnapali Vohra, Priya Taylor, Steve M. Vo-Dinh, Tuan Sci Rep Article Direct detection of genetic biomarkers in body fluid lysate without target amplification will revolutionize nucleic acid-based diagnostics. However, the low concentration of target sequences makes this goal challenging. We report a method for direct detection of pathogen RNA in blood lysate using a bioassay using surface-enhanced Raman spectroscopy (SERS)-based detection integrated in a “lab-in-a-stick” portable device. Two levels of signal enhancement were employed to achieve the sensitivity required for direct detection. Each target sequence was tagged with an ultrabright SERS-encoded nanorattle with ultrahigh SERS signals, and these tagged target sequences were concentrated into a focused spot for detection using hybridization sandwiches with magnetic microbeads. Furthermore, the washing process was automated by integration into a “lab-in-a-stick” portable device. We could directly detect synthetic target with a limit of detection of 200 fM. More importantly, we detected plasmodium falciparum malaria parasite RNA directly in infected red blood cells lysate. To our knowledge, this is the first report of SERS-based direct detection of pathogen nucleic acid in blood lysate without nucleic acid extraction or target amplification. The results show the potential of our integrated bioassay for field use and point-of-care diagnostics. Nature Publishing Group UK 2018-03-06 /pmc/articles/PMC5840326/ /pubmed/29511216 http://dx.doi.org/10.1038/s41598-018-21615-3 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Ngo, Hoan T. Freedman, Elizabeth Odion, Ren Abelard Strobbia, Pietro De Silva Indrasekara, Agampodi Swarnapali Vohra, Priya Taylor, Steve M. Vo-Dinh, Tuan Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles |
title | Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles |
title_full | Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles |
title_fullStr | Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles |
title_full_unstemmed | Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles |
title_short | Direct Detection of Unamplified Pathogen RNA in Blood Lysate using an Integrated Lab-in-a-Stick Device and Ultrabright SERS Nanorattles |
title_sort | direct detection of unamplified pathogen rna in blood lysate using an integrated lab-in-a-stick device and ultrabright sers nanorattles |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5840326/ https://www.ncbi.nlm.nih.gov/pubmed/29511216 http://dx.doi.org/10.1038/s41598-018-21615-3 |
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