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Research Article

Super-assembly of integrated gold magnetic assay with loop-mediated isothermal amplification for point-of-care testing

Jianping Liang1,§, Jie Zeng3,§, Xiaojuan Huang1, Tengteng Zhu1, Yonglong Gong1, Chen Dong1, Xiangrong Wang1,5, Lingzhi Zhao1, Lei Xie3, Kang Liang6, Qiongxiang Tan1, Yali Cui1,2( ), Biao Kong3( ), Wenli Hui1,4( )
The College of life science, Northwest University, Xi’an 710069, China
Shaanxi Provincial Engineering Research Center for Nano-Biomedical Detection, Xi’an 710077, China
Department of Chemistry, Shanghai Key Lab of Molecular Catalysis and Innovative Materials, Collaborative Innovation Center of Chemistry for Energy Materials, Fudan University, Shanghai 200438, China
Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University, Xi’an 710069, China
Provincial Key Laboratory of Biotechnology of Shaanxi Province, Northwest University, Xi’an 710069, China
School of Chemical Engineering, Graduate School of Biomedical Engineering, and Australian Centre for NanoMedicine, University of New South Wales, NSW 2052, Australia

§ Jianping Liang and Jie Zeng contributed equally to this work.

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Graphical Abstract

The schematic illustration displays the over-all process of surface modification and molecular detection of gold magnetic nanoparticles. The anti-digoxin antibody labeled on the modified gold magnetic nanoparticles and loop-mediated isothermal amplification (LAMP) double-labeled products formed by biotin and digoxin labeled on primers interact with the lateral flow assay (LFA) system and present visual bands.

Abstract

With the increasing global threat of various diseases and infections, it is essential to develop a fast, low-cost, and easy-to-use point-of-care testing (POCT) system for inspections at all levels of medical institutions and self-examination at home. In this work, gold magnetic nanoparticles (GMNPs) are used as the key material, and a rapid visual detection method is designed through integrating loop-mediated isothermal amplification (LAMP) and lateral flow assay (LFA) biosensor for detecting a variety of analytes which includes whole blood, buccal swabs, and DNA. It is worth to note that the proposed method does not need DNA extraction. Furthermore, uracil DNA glycosylase (UDG) is employed to eliminate carrier contamination for preventing false positive results. The whole detection process can be finished within 25 min. The accuracy of detection is measured by assessing the polymorphisms of the methylenetetrahydrofolate reductase (MTHFR) C677T. The detection limit of the newly developed extraction-free detection system for MTHFR C677T is 0.16 ng/μL. A preliminary clinical study of the proposed method is carried out by analyzing 600 clinical samples (including 200 whole blood samples, 100 buccal swabs, and 300 genomic DNA samples). The results indicate that the proposed method is 100% consistent with the sequencing results which provides a new choice for POCT and shows a broad application prospect in all levels of medical clinics and at home.

Keywords

gold magnetic nanoparticlesloop-mediated isothermal amplificationlateral flow assay systemfree extractionsingle-nucleotide polymorphisms genotyping

Electronic Supplementary Material

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Nano Research
Volume 16 Issue 1,
January 2023
Pages 1242-1251
DOI: 10.1007/s12274-022-4692-9
Cite this article:
Liang J, Zeng J, Huang X, et al. Super-assembly of integrated gold magnetic assay with loop-mediated isothermal amplification for point-of-care testing. Nano Research, 2023, 16(1): 1242-1251. https://doi.org/10.1007/s12274-022-4692-9
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Received: 25 May 2022
Revised: 21 June 2022
Accepted: 21 June 2022
Published: 04 August 2022
© Tsinghua University Press 2022
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