Microfluidic Encapsulation of DNAs in Liquid Beads for Digital Loop-Mediated Isothermal Amplification

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Galogahi, Fariba Malekpour
Strachan, Simon
Yadav, Ajeet Singh
Stratton, Helen
Nguyen, Nam‐Trung
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2024
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Abstract

Digital nucleic acid analysis has emerged as a prominent tool for the detection and absolute quantification of diverse pathogens. Digital loop-mediated isothermal amplification (dLAMP) offers highly sensitive, specific, time-efficient, and cost-effective nucleic acid amplification. However, existing dLAMP techniques face challenges such as droplet merging, reliance on surfactants, restricted partition capacities, and the potential for sample loss during heating. Herein, these issues are addressed by introducing liquid beads for sample partitioning. Compared to microwells, our approach overcomes the limitations of chamber dimensions, enabling the analysis of an unlimited number of digitized targets. Furthermore, our novel approach effectively addresses sample loss and merging during thermal processing and eliminates the need for surfactants. Accurate and reproducible the quantitative detection of the gene cluster XALB1 of leaf scald disease is conducted using dLAMP based on liquid beads to verify its availability. The results demonstrate a high correlation between target concentration and positive signals, indicating the robust performance of our technique. A comparative analysis is then performed between dLAMP using liquid beads and using single droplets. Benchmarking these two techniques highlights the effectiveness of our innovative technique in overcoming existing challenges in dLAMP.

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Advanced NanoBiomed Research

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© 2024 The Author(s). Advanced NanoBiomed Research published by Wiley-VCH GmbH. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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This publication has been entered in Griffith Research Online as an advance online version.

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Nanobiotechnology

Nanotechnology

Genetics

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Galogahi, FM; Strachan, S; Yadav, AS; Stratton, H; Nguyen, N, Microfluidic Encapsulation of DNAs in Liquid Beads for Digital Loop-Mediated Isothermal Amplification, Advanced NanoBiomed Research, 2024, pp. 2400044

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