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一种用于DNA样品制备、等温扩增和无标记电化学检测的集成式电子管帽。

An integrated E-Tube cap for sample preparation, isothermal amplification and label-free electrochemical detection of DNA.

作者信息

Xu Zhiheng, Yin Kun, Ding Xiong, Li Ziyue, Sun Xuanhao, Li Baikun, Lalla Rajesh V, Gross Robert, Liu Changchun

机构信息

Department of Biomedical Engineering, University of Connecticut Health Center, Farmington, CT, 06030, USA.

Department of Physiology and Neurobiology, University of Connecticut, Storrs, CT, 06269, USA.

出版信息

Biosens Bioelectron. 2021 Aug 15;186:113306. doi: 10.1016/j.bios.2021.113306. Epub 2021 May 7.


DOI:10.1016/j.bios.2021.113306
PMID:33991846
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8572321/
Abstract

A simple, disposable, and integrated electronic-tube cap (E-tube cap) for DNA detection at the point-of-care was designed, fabricated, and tested. The E-tube cap contains a 3D printed electrode substrate for DNA extraction and label-free pH sensing detection. One Flinders Technology Associates (Whatman FTA) membrane was incorporated into the 3D printed electrode substrate for the isolation, concentration, and purification of DNA. The E-tube cap with captured DNA by the membrane was inserted directly into a reaction tube for loop-mediated isothermal amplification (LAMP). The isothermal amplification process was monitored in real-time by a 3D printed electrochemical electrode coated with pH-sensitive material (carbon/iridium oxide layer). The pH sensing electrode showed an excellent linear response within the pH range of 6-9 with a slope of -31.32 ± 0.5 mV/pH at room temperature. The utility of the integrated E-tube cap was demonstrated by detecting the presence of lambda DNA spiked in saliva samples with a sensitivity of 100 copies per mL sample within 30 min. Such a simple, rapid, and affordable diagnostic device is particularly suitable for point-of-care molecular diagnostics of infectious diseases.

摘要

设计、制造并测试了一种用于即时检测DNA的简单、一次性且集成的电子管帽(E管帽)。该E管帽包含一个用于DNA提取和无标记pH传感检测的3D打印电极基板。将一张弗林德斯技术协会(Whatman FTA)膜并入3D打印电极基板中,用于DNA的分离、浓缩和纯化。将通过膜捕获了DNA的E管帽直接插入反应管中进行环介导等温扩增(LAMP)。通过涂有pH敏感材料(碳/氧化铱层)的3D打印电化学电极实时监测等温扩增过程。在室温下,pH传感电极在6-9的pH范围内显示出优异的线性响应,斜率为-31.32±0.5 mV/pH。通过检测唾液样本中添加的λDNA,在30分钟内灵敏度达到每毫升样本100个拷贝,证明了集成E管帽的实用性。这种简单、快速且经济实惠的诊断设备特别适用于传染病的即时分子诊断。

相似文献

[1]
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引用本文的文献

[1]
Manually-Operated, Slider Cassette for Multiplexed Molecular Detection at the Point of Care.

Sens Actuators B Chem. 2022-10-15

[2]
Chemical Trends in Sample Preparation for Nucleic Acid Amplification Testing (NAAT): A Review.

Biosensors (Basel). 2023-11-10

[3]
Real-time monitoring of isothermal nucleic acid amplification on a smartphone by using a portable electrochemical device for home-testing of SARS-CoV-2.

Anal Chim Acta. 2022-10-9

本文引用的文献

[1]
ZIF-8-Assisted NaYF4:Yb,Tm@ZnO Converter with Exonuclease III-Powered DNA Walker for Near-Infrared Light Responsive Biosensor.

Anal Chem. 2019-12-9

[2]
Palindromic Molecular Beacon Based Z-Scheme BiOCl-Au-CdS Photoelectrochemical Biodetection.

Anal Chem. 2019-1-16

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Fully 3D printed integrated reactor array for point-of-care molecular diagnostics.

Biosens Bioelectron. 2018-3-10

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Ticks Tick Borne Dis. 2017-6

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An integrated rotary microfluidic system with DNA extraction, loop-mediated isothermal amplification, and lateral flow strip based detection for point-of-care pathogen diagnostics.

Biosens Bioelectron. 2016-12-14

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Target-regulated proximity hybridization with three-way DNA junction for in situ enhanced electronic detection of marine biotoxin based on isothermal cycling signal amplification strategy.

Biosens Bioelectron. 2015-2-26

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Using the ubiquitous pH meter combined with a loop mediated isothermal amplification method for facile and sensitive detection of Nosema bombycis genomic DNA PTP1.

Chem Commun (Camb). 2014-12-28

[8]
A real-time microfluidic multiplex electrochemical loop-mediated isothermal amplification chip for differentiating bacteria.

Biosens Bioelectron. 2014-4-8

[9]
Real-time fluorescence loop mediated isothermal amplification for the detection of Acinetobacter baumannii.

PLoS One. 2013-7-2

[10]
Simultaneous DNA amplification and detection using a pH-sensing semiconductor system.

Nat Methods. 2013-6-9

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