Multi-frequency impedance sensing for detection and sizing of DNA fragments

Abstract Electronic biosensors for DNA detection typically utilize immobilized oligonucleotide probes on a signal transducer, which outputs an electronic signal when target molecules bind to probes. However, limitation in probe selectivity and variable levels of non-target material in complex biolog...

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Autores principales:	Jianye Sui, Neeru Gandotra, Pengfei Xie, Zhongtian Lin, Curt Scharfe, Mehdi Javanmard
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6fdf1e2ab58b436883cd2570cd574cc3
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spelling	oai:doaj.org-article:6fdf1e2ab58b436883cd2570cd574cc32021-12-02T13:24:14ZMulti-frequency impedance sensing for detection and sizing of DNA fragments10.1038/s41598-021-85755-92045-2322https://doaj.org/article/6fdf1e2ab58b436883cd2570cd574cc32021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-85755-9https://doaj.org/toc/2045-2322Abstract Electronic biosensors for DNA detection typically utilize immobilized oligonucleotide probes on a signal transducer, which outputs an electronic signal when target molecules bind to probes. However, limitation in probe selectivity and variable levels of non-target material in complex biological samples can lead to nonspecific binding and reduced sensitivity. Here we introduce the integration of 2.8 μm paramagnetic beads with DNA fragments. We apply a custom-made microfluidic chip to detect DNA molecules bound to beads by measuring Impedance Peak Response (IPR) at multiple frequencies. Technical and analytical performance was evaluated using beads containing purified Polymerase Chain Reaction (PCR) products of different lengths (157, 300, 613 bp) with DNA concentration ranging from 0.039 amol to 7.8 fmol. Multi-frequency IPR correlated positively with DNA amounts and was used to calculate a DNA quantification score. The minimum DNA amount of a 300 bp fragment coupled on beads that could be robustly detected was 0.0039 fmol (1.54 fg or 4750 copies/bead). Additionally, our approach allowed distinguishing beads with similar molar concentration DNA fragments of different lengths. Using this impedance sensor, purified PCR products could be analyzed within ten minutes to determine DNA fragment length and quantity based on comparison to a known DNA standard.Jianye SuiNeeru GandotraPengfei XieZhongtian LinCurt ScharfeMehdi JavanmardNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jianye Sui Neeru Gandotra Pengfei Xie Zhongtian Lin Curt Scharfe Mehdi Javanmard Multi-frequency impedance sensing for detection and sizing of DNA fragments
description	Abstract Electronic biosensors for DNA detection typically utilize immobilized oligonucleotide probes on a signal transducer, which outputs an electronic signal when target molecules bind to probes. However, limitation in probe selectivity and variable levels of non-target material in complex biological samples can lead to nonspecific binding and reduced sensitivity. Here we introduce the integration of 2.8 μm paramagnetic beads with DNA fragments. We apply a custom-made microfluidic chip to detect DNA molecules bound to beads by measuring Impedance Peak Response (IPR) at multiple frequencies. Technical and analytical performance was evaluated using beads containing purified Polymerase Chain Reaction (PCR) products of different lengths (157, 300, 613 bp) with DNA concentration ranging from 0.039 amol to 7.8 fmol. Multi-frequency IPR correlated positively with DNA amounts and was used to calculate a DNA quantification score. The minimum DNA amount of a 300 bp fragment coupled on beads that could be robustly detected was 0.0039 fmol (1.54 fg or 4750 copies/bead). Additionally, our approach allowed distinguishing beads with similar molar concentration DNA fragments of different lengths. Using this impedance sensor, purified PCR products could be analyzed within ten minutes to determine DNA fragment length and quantity based on comparison to a known DNA standard.
format	article
author	Jianye Sui Neeru Gandotra Pengfei Xie Zhongtian Lin Curt Scharfe Mehdi Javanmard
author_facet	Jianye Sui Neeru Gandotra Pengfei Xie Zhongtian Lin Curt Scharfe Mehdi Javanmard
author_sort	Jianye Sui
title	Multi-frequency impedance sensing for detection and sizing of DNA fragments
title_short	Multi-frequency impedance sensing for detection and sizing of DNA fragments
title_full	Multi-frequency impedance sensing for detection and sizing of DNA fragments
title_fullStr	Multi-frequency impedance sensing for detection and sizing of DNA fragments
title_full_unstemmed	Multi-frequency impedance sensing for detection and sizing of DNA fragments
title_sort	multi-frequency impedance sensing for detection and sizing of dna fragments
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/6fdf1e2ab58b436883cd2570cd574cc3
work_keys_str_mv	AT jianyesui multifrequencyimpedancesensingfordetectionandsizingofdnafragments AT neerugandotra multifrequencyimpedancesensingfordetectionandsizingofdnafragments AT pengfeixie multifrequencyimpedancesensingfordetectionandsizingofdnafragments AT zhongtianlin multifrequencyimpedancesensingfordetectionandsizingofdnafragments AT curtscharfe multifrequencyimpedancesensingfordetectionandsizingofdnafragments AT mehdijavanmard multifrequencyimpedancesensingfordetectionandsizingofdnafragments
_version_	1718393099715084288

Multi-frequency impedance sensing for detection and sizing of DNA fragments

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