Gold nanoparticles – an optical biosensor for RNA quantification for cancer and neurologic disorders diagnosis

Gold nanoparticles – an optical biosensor for RNA quantification for cancer and neurologic disorders diagnosis

Sherif M Shawky,1–3,* Ahmed M Awad,1,4,* Arwa A Abugable,1,2,* Sherif F El-Khamisy1,2 1Center for Genomics, Helmy Institute for Medical Sciences, Zewail City of Science and Technology, Giza, Egypt; 2Krebs Institute, Department of Molecular Biology and Biotechnology, Firth Court,...

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Autores principales: Shawky SM, Awad AM, Abugable AA, El-Khamisy SF
Formato: article
Lenguaje:EN
Publicado: Dove Medical Press 2018
Materias:
Gold Aggregating Gold (GAG)
TOP1
TDP2
Magnetic nanoparticles
RNA extraction
Medicine (General)
R5-920
Acceso en línea:https://doaj.org/article/3eaf74705e6c44d6b94961368b76c319
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spelling oai:doaj.org-article:3eaf74705e6c44d6b94961368b76c3192021-12-02T03:49:59ZGold nanoparticles – an optical biosensor for RNA quantification for cancer and neurologic disorders diagnosis1178-2013https://doaj.org/article/3eaf74705e6c44d6b94961368b76c3192018-11-01T00:00:00Zhttps://www.dovepress.com/gold-nanoparticles-an-optical-biosensor-for-rna-quantification-for-can-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Sherif M Shawky,1–3,* Ahmed M Awad,1,4,* Arwa A Abugable,1,2,* Sherif F El-Khamisy1,2 1Center for Genomics, Helmy Institute for Medical Sciences, Zewail City of Science and Technology, Giza, Egypt; 2Krebs Institute, Department of Molecular Biology and Biotechnology, Firth Court, University of Sheffield, Sheffield, UK; 3Biochemistry Department, Faculty of Pharmacy, Misr University for Science and Technology, Giza, Egypt; 4Department of Molecular Biology, Genetic Engineering and Biotechnology Research Institute, University of Sadat City, Sadat City, Egypt *These authors contributed equally to this work Purpose: The objective of this study is to develop a facile tool for the absolute detection and quantification of nucleic acid transcripts, using a gold nanoparticle-based optical biosensor. Topoisomerase 1 (TOP1) and tyrosyl DNA phosphodiesterase 2 (TDP2) were among the nucleic acid transcripts of choice due to their role as genomic instability biomarkers and their implication in various cancers and neurologic disorders. This opens the door to develop a simple tool that can be used for diagnosing and monitoring treatment response for such diseases, overcoming the requirements for high cost, time, and complexity of the existing technologies for the absolute quantification of transcripts of interest. Materials and methods: The TOP1 and TDP2 mRNA transcripts were first captured specifically using magnetic nanoparticles that were functionalized with TOP1- and TDP2-specific probes, respectively. The captured mRNA was then directly detected and quantified using the gold aggregating gold (GAG) assay, without the need for amplification as in existing technologies used for the quantification of transcripts. Results: A linear correlation exists between the GAG assay and the qPCR for the quantification of the TOP1 and TDP2 mRNA transcripts (101–104 copies). The detection limit of the GAG assay in mRNA quantification was up to 10 copies per reaction. Wild-type and TDP2-deficient cell lines confirmed the assay specificity and reproducibility in distinguishing between different transcripts. Conclusion: The GAG assay can be utilized as an inexpensive, rapid, simple, and sensitive tool for the absolute quantification of RNA for different applications, instead of the laborious, expensive, and sophisticated real-time PCR. Keywords: gold aggregating gold, TOP1, TDP2, magnetic nanoparticles, RNA extraction, genomic instability, neurodegenerative diseases, tumor markersShawky SMAwad AMAbugable AAEl-Khamisy SFDove Medical PressarticleGold Aggregating Gold (GAG)TOP1TDP2Magnetic nanoparticlesRNA extractionMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 13, Pp 8137-8151 (2018)
institution DOAJ
collection DOAJ
language EN
topic Gold Aggregating Gold (GAG)
TOP1
TDP2
Magnetic nanoparticles
RNA extraction
Medicine (General)
R5-920
spellingShingle Gold Aggregating Gold (GAG)
TOP1
TDP2
Magnetic nanoparticles
RNA extraction
Medicine (General)
R5-920
Shawky SM
Awad AM
Abugable AA
El-Khamisy SF
Gold nanoparticles – an optical biosensor for RNA quantification for cancer and neurologic disorders diagnosis
description Sherif M Shawky,1–3,* Ahmed M Awad,1,4,* Arwa A Abugable,1,2,* Sherif F El-Khamisy1,2 1Center for Genomics, Helmy Institute for Medical Sciences, Zewail City of Science and Technology, Giza, Egypt; 2Krebs Institute, Department of Molecular Biology and Biotechnology, Firth Court, University of Sheffield, Sheffield, UK; 3Biochemistry Department, Faculty of Pharmacy, Misr University for Science and Technology, Giza, Egypt; 4Department of Molecular Biology, Genetic Engineering and Biotechnology Research Institute, University of Sadat City, Sadat City, Egypt *These authors contributed equally to this work Purpose: The objective of this study is to develop a facile tool for the absolute detection and quantification of nucleic acid transcripts, using a gold nanoparticle-based optical biosensor. Topoisomerase 1 (TOP1) and tyrosyl DNA phosphodiesterase 2 (TDP2) were among the nucleic acid transcripts of choice due to their role as genomic instability biomarkers and their implication in various cancers and neurologic disorders. This opens the door to develop a simple tool that can be used for diagnosing and monitoring treatment response for such diseases, overcoming the requirements for high cost, time, and complexity of the existing technologies for the absolute quantification of transcripts of interest. Materials and methods: The TOP1 and TDP2 mRNA transcripts were first captured specifically using magnetic nanoparticles that were functionalized with TOP1- and TDP2-specific probes, respectively. The captured mRNA was then directly detected and quantified using the gold aggregating gold (GAG) assay, without the need for amplification as in existing technologies used for the quantification of transcripts. Results: A linear correlation exists between the GAG assay and the qPCR for the quantification of the TOP1 and TDP2 mRNA transcripts (101–104 copies). The detection limit of the GAG assay in mRNA quantification was up to 10 copies per reaction. Wild-type and TDP2-deficient cell lines confirmed the assay specificity and reproducibility in distinguishing between different transcripts. Conclusion: The GAG assay can be utilized as an inexpensive, rapid, simple, and sensitive tool for the absolute quantification of RNA for different applications, instead of the laborious, expensive, and sophisticated real-time PCR. Keywords: gold aggregating gold, TOP1, TDP2, magnetic nanoparticles, RNA extraction, genomic instability, neurodegenerative diseases, tumor markers
format article
author Shawky SM
Awad AM
Abugable AA
El-Khamisy SF
author_facet Shawky SM
Awad AM
Abugable AA
El-Khamisy SF
author_sort Shawky SM
title Gold nanoparticles – an optical biosensor for RNA quantification for cancer and neurologic disorders diagnosis
title_short Gold nanoparticles – an optical biosensor for RNA quantification for cancer and neurologic disorders diagnosis
title_full Gold nanoparticles – an optical biosensor for RNA quantification for cancer and neurologic disorders diagnosis
title_fullStr Gold nanoparticles – an optical biosensor for RNA quantification for cancer and neurologic disorders diagnosis
title_full_unstemmed Gold nanoparticles – an optical biosensor for RNA quantification for cancer and neurologic disorders diagnosis
title_sort gold nanoparticles – an optical biosensor for rna quantification for cancer and neurologic disorders diagnosis
publisher Dove Medical Press
publishDate 2018
url https://doaj.org/article/3eaf74705e6c44d6b94961368b76c319
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AT abugableaa goldnanoparticlesndashanopticalbiosensorforrnaquantificationforcancerandneurologicdisordersdiagnosis
AT elkhamisysf goldnanoparticlesndashanopticalbiosensorforrnaquantificationforcancerandneurologicdisordersdiagnosis
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