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iso-μmGene: an isothermal amplification-based portable microfluidic system for simple, reliable and flexibly multiplexed genetic identification and quantification

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Abstract

Simple, reliable and flexibly multiplexed genetic identification and quantification of microbial pathogens is in urgent need for early disease diagnosis and timely treatment. This study presented an isothermal amplification-based portable microfluidic system (iso-μmGene) with features of multi-well chips for convenient filling and reliable sealing, flexible detection throughput, and a stand-alone and well-performing point of care (POC) genetic testing device. Using disposable chips with two kinds of reaction wells (eighteen and ten wells) and a device prototype with independent four chip holders, the iso-μmGene enables on-demand analysis of different target genes in one sample per chip and one to four samples (chips) per run, requiring only a single pipetting step for dispensing per chip with dehydrated primers. To completely seal the loop-mediated isothermal amplification (LAMP) reaction system to minimize the risk of amplicon escape, a dedicated plastic shell is used to assemble the array-type chip and reliably close its openings. Meanwhile, to enhance the precision for flexibly multiplexed detection and decrease the size and cost of the device, we designed a thermoelectric cooler (TEC)-based temperature-control module including two separate units and a CCD-based fluorescence imaging module containing a linear translation stage for real-time LAMP assay. This work demonstrated applications for the parallel detection of 2–2000 CFU (colony forming units) per reaction well with good intra- and inter-chip reproducibility using the crude lysates of two aquaculture pathogens Edwardsiella tarda and Vibrio harveyi. Overall, the iso-μmGene presented here possesses both a sophisticated instrument's functionality and performance and POC device's portability and cost.

Graphical abstract: iso-μmGene: an isothermal amplification-based portable microfluidic system for simple, reliable and flexibly multiplexed genetic identification and quantification

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Supplementary files

Article information


Submitted
20 Mar 2020
Accepted
30 Apr 2020
First published
01 May 2020

Analyst, 2020, Advance Article
Article type
Paper

iso-μmGene: an isothermal amplification-based portable microfluidic system for simple, reliable and flexibly multiplexed genetic identification and quantification

R. Zhong, S. Liu, G. Zhang, M. Wang and Y. Sun, Analyst, 2020, Advance Article , DOI: 10.1039/D0AN00560F

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