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Sensitive, simultaneous quantitation of two unlabeled DNA targets using a magnetic nanoparticle-enzyme sandwich assay

We report herein the development of a simple, sensitive colorimetric magnetic nanoparticle (MNP)-enzyme-based DNA sandwich assay that is suitable for simultaneous, label-free quantitation of two DNA targets down to 50 fM level. It can also effectively discriminate single-nucleotide polymorphisms (SN... Full description

Journal Title: Analytical chemistry 01 October 2013, Vol.85(19), pp.9238-44
Main Author: Zhang, Yue
Other Authors: Pilapong, Chalermchai , Guo, Yuan , Ling, Zhenlian , Cespedes, Oscar , Quirke, Philip , Zhou, Dejian
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: E-ISSN: 1520-6882 ; PMID: 23971744 Version:1 ; DOI: 10.1021/ac402081u
Link: http://pubmed.gov/23971744
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recordid: medline23971744
title: Sensitive, simultaneous quantitation of two unlabeled DNA targets using a magnetic nanoparticle-enzyme sandwich assay
format: Article
creator:
  • Zhang, Yue
  • Pilapong, Chalermchai
  • Guo, Yuan
  • Ling, Zhenlian
  • Cespedes, Oscar
  • Quirke, Philip
  • Zhou, Dejian
subjects:
  • Biosensing Techniques -- Methods
  • DNA Probes -- Chemistry
  • DNA, Neoplasm -- Analysis
  • Enzymes -- Metabolism
  • Magnetite Nanoparticles -- Chemistry
ispartof: Analytical chemistry, 01 October 2013, Vol.85(19), pp.9238-44
description: We report herein the development of a simple, sensitive colorimetric magnetic nanoparticle (MNP)-enzyme-based DNA sandwich assay that is suitable for simultaneous, label-free quantitation of two DNA targets down to 50 fM level. It can also effectively discriminate single-nucleotide polymorphisms (SNPs) in genes associated with human cancers (KRAS codon 12/13 SNPs). This assay uses a pair of specific DNA probes, one being covalently conjugated to an MNP for target capture and the other being linked to an enzyme for signal amplification, to sandwich a DNA target, allowing for convenient magnetic separation and subsequent efficient enzymatic signal amplification for high sensitivity. Careful optimization of the MNP surfaces and assay conditions greatly reduced the background, allowing for sensitive, specific detection of as little as 5 amol (50 fM in 100 μL) of target DNA. Moreover, this sensor is robust, it can effectively discriminate cancer-specific SNPs against the wild-type noncancer...
language: eng
source:
identifier: E-ISSN: 1520-6882 ; PMID: 23971744 Version:1 ; DOI: 10.1021/ac402081u
fulltext: fulltext
issn:
  • 15206882
  • 1520-6882
url: Link


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titleSensitive, simultaneous quantitation of two unlabeled DNA targets using a magnetic nanoparticle-enzyme sandwich assay
creatorZhang, Yue ; Pilapong, Chalermchai ; Guo, Yuan ; Ling, Zhenlian ; Cespedes, Oscar ; Quirke, Philip ; Zhou, Dejian
ispartofAnalytical chemistry, 01 October 2013, Vol.85(19), pp.9238-44
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subjectBiosensing Techniques -- Methods ; DNA Probes -- Chemistry ; DNA, Neoplasm -- Analysis ; Enzymes -- Metabolism ; Magnetite Nanoparticles -- Chemistry
descriptionWe report herein the development of a simple, sensitive colorimetric magnetic nanoparticle (MNP)-enzyme-based DNA sandwich assay that is suitable for simultaneous, label-free quantitation of two DNA targets down to 50 fM level. It can also effectively discriminate single-nucleotide polymorphisms (SNPs) in genes associated with human cancers (KRAS codon 12/13 SNPs). This assay uses a pair of specific DNA probes, one being covalently conjugated to an MNP for target capture and the other being linked to an enzyme for signal amplification, to sandwich a DNA target, allowing for convenient magnetic separation and subsequent efficient enzymatic signal amplification for high sensitivity. Careful optimization of the MNP surfaces and assay conditions greatly reduced the background, allowing for sensitive, specific detection of as little as 5 amol (50 fM in 100 μL) of target DNA. Moreover, this sensor is robust, it can effectively discriminate cancer-specific SNPs against the wild-type noncancer...
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abstractWe report herein the development of a simple, sensitive colorimetric magnetic nanoparticle (MNP)-enzyme-based DNA sandwich assay that is suitable for simultaneous, label-free quantitation of two DNA targets down to 50 fM level. It can also effectively discriminate single-nucleotide polymorphisms (SNPs) in genes associated with human cancers (KRAS codon 12/13 SNPs). This assay uses a pair of specific DNA probes, one being covalently conjugated to an MNP for target capture and the other being linked to an enzyme for signal amplification, to sandwich a DNA target, allowing for convenient magnetic separation and subsequent efficient enzymatic signal amplification for high sensitivity. Careful optimization of the MNP surfaces and assay conditions greatly reduced the background, allowing for sensitive, specific detection of as little as 5 amol (50 fM in 100 μL) of target DNA. Moreover, this sensor is robust, it can effectively discriminate cancer-specific SNPs against the wild-type noncancer...
doi10.1021/ac402081u
pmid23971744
date2013-10-01