An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA Amplification
The sensitive quantification of low-abundance nucleic acids holds importance for a range of clinical applications and biological studies. In this study, we describe a facile microfluidic chip for absolute DNA quantifications based on the digital loop-mediated isothermal amplification (digital LAMP)...
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doaj-e9d0e529d00f49bb8457cea8500bfe4a2020-11-25T01:30:16ZengMDPI AGMicromachines2072-666X2020-02-0111217710.3390/mi11020177mi11020177An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA AmplificationZengming Zhang0Shuhao Zhao1Fei Hu2Guangpu Yang3Juan Li4Hui Tian5Niancai Peng6State Key Laboratory for Manufacturing Systems Engineering, Xi’an Jiaotong University, Xi’an, 710054, Shaanxi, ChinaState Key Laboratory for Manufacturing Systems Engineering, Xi’an Jiaotong University, Xi’an, 710054, Shaanxi, ChinaState Key Laboratory for Manufacturing Systems Engineering, Xi’an Jiaotong University, Xi’an, 710054, Shaanxi, ChinaState Key Laboratory for Manufacturing Systems Engineering, Xi’an Jiaotong University, Xi’an, 710054, Shaanxi, ChinaState Key Laboratory for Manufacturing Systems Engineering, Xi’an Jiaotong University, Xi’an, 710054, Shaanxi, ChinaState Key Laboratory for Manufacturing Systems Engineering, Xi’an Jiaotong University, Xi’an, 710054, Shaanxi, ChinaState Key Laboratory for Manufacturing Systems Engineering, Xi’an Jiaotong University, Xi’an, 710054, Shaanxi, ChinaThe sensitive quantification of low-abundance nucleic acids holds importance for a range of clinical applications and biological studies. In this study, we describe a facile microfluidic chip for absolute DNA quantifications based on the digital loop-mediated isothermal amplification (digital LAMP) method. This microfluidic chip integrates a cross-flow channel for droplet generation with a micro-cavity for droplet tiling. DNA templates in the LAMP reagent were divided into ~20,000 water-in-oil droplets at the cross-flow channel. The droplets were then tiled in the micro-cavity for isothermal amplification and fluorescent detection. Different from the existing polydimethylsiloxane (PDMS) microfluidic chips, this study incorporates gold nanoparticles (AuNPs) into PDMS substrate through silica coating and dodecanol modification. The digital LAMP chip prepared by AuNPs-PDMS combines the benefits of the microstructure manufacturing performance of PDMS with the light-to-heat conversion advantages of AuNPs. Upon illumination with a near infrared (NIR) LED, the droplets were stably and efficiently heated by the AuNPs in PDMS. We further introduce an integrated device with a NIR heating unit and a fluorescent detection unit. The system could detect HBV (hepatitis B virus)-DNA at a concentration of 1 × 10<sup>1</sup> to 1 × 10<sup>4</sup> copies/μL. The LED-driven digital LAMP chip and the integrated device; therefore, demonstrate high accuracy and excellent performance for the absolute quantification of low-abundance nucleic acids, showing the advantages of integration, miniaturization, cost, and power consumption.https://www.mdpi.com/2072-666X/11/2/177digital lampgold nanoparticlesintegrated devicedna quantification |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Zengming Zhang Shuhao Zhao Fei Hu Guangpu Yang Juan Li Hui Tian Niancai Peng |
spellingShingle |
Zengming Zhang Shuhao Zhao Fei Hu Guangpu Yang Juan Li Hui Tian Niancai Peng An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA Amplification Micromachines digital lamp gold nanoparticles integrated device dna quantification |
author_facet |
Zengming Zhang Shuhao Zhao Fei Hu Guangpu Yang Juan Li Hui Tian Niancai Peng |
author_sort |
Zengming Zhang |
title |
An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA Amplification |
title_short |
An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA Amplification |
title_full |
An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA Amplification |
title_fullStr |
An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA Amplification |
title_full_unstemmed |
An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA Amplification |
title_sort |
led-driven aunps-pdms microfluidic chip and integrated device for the detection of digital loop-mediated isothermal dna amplification |
publisher |
MDPI AG |
series |
Micromachines |
issn |
2072-666X |
publishDate |
2020-02-01 |
description |
The sensitive quantification of low-abundance nucleic acids holds importance for a range of clinical applications and biological studies. In this study, we describe a facile microfluidic chip for absolute DNA quantifications based on the digital loop-mediated isothermal amplification (digital LAMP) method. This microfluidic chip integrates a cross-flow channel for droplet generation with a micro-cavity for droplet tiling. DNA templates in the LAMP reagent were divided into ~20,000 water-in-oil droplets at the cross-flow channel. The droplets were then tiled in the micro-cavity for isothermal amplification and fluorescent detection. Different from the existing polydimethylsiloxane (PDMS) microfluidic chips, this study incorporates gold nanoparticles (AuNPs) into PDMS substrate through silica coating and dodecanol modification. The digital LAMP chip prepared by AuNPs-PDMS combines the benefits of the microstructure manufacturing performance of PDMS with the light-to-heat conversion advantages of AuNPs. Upon illumination with a near infrared (NIR) LED, the droplets were stably and efficiently heated by the AuNPs in PDMS. We further introduce an integrated device with a NIR heating unit and a fluorescent detection unit. The system could detect HBV (hepatitis B virus)-DNA at a concentration of 1 × 10<sup>1</sup> to 1 × 10<sup>4</sup> copies/μL. The LED-driven digital LAMP chip and the integrated device; therefore, demonstrate high accuracy and excellent performance for the absolute quantification of low-abundance nucleic acids, showing the advantages of integration, miniaturization, cost, and power consumption. |
topic |
digital lamp gold nanoparticles integrated device dna quantification |
url |
https://www.mdpi.com/2072-666X/11/2/177 |
work_keys_str_mv |
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