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Microfluidic Biosensors for Point-of-Care Nucleic Acid Amplification Tests

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A special issue of Biosensors (ISSN 2079-6374). This special issue belongs to the section "Biosensors and Healthcare".

Deadline for manuscript submissions: closed (31 October 2023) | Viewed by 11551

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Special Issue Editors

Dr. Kieu The Loan Trinh

E-Mail Website
Guest Editor

Bionano Applications Research Center, Gachon University, Seongnam-si 13120, Republic of Korea
Interests: lab-on-a-chip; system integration; nucleic acid detection
Special Issues, Collections and Topics in MDPI journals

Prof. Dr. Xianbo Lu

E-Mail Website
Guest Editor

Dalian Institute of Chemical Physics, Chinese Academy of Science, Dalian 116023, China
Interests: electrochemical sensors and biosensors; enzyme biosensors; nucleic acid sensors; aptasensor; screen-printed electrodes; microfluidic paper based analytical device (mPAD); nanosensing materials; nanoenzyme; toxic effect evaluation

Special Issue Information

Dear Colleagues,

We are pleased to invite contributions to this Special Issue covering relevant microfluidic-based technologies for molecular sensing and diagnosis, such as nucleic acid amplification tests. This Special Issue aims to gather original research papers and comprehensive reviews revealing novel research, fabrication methods, and applications as well as the challenges and prospects in developing microfluidics for improved biosensing and diagnostics in healthcare, food safety, and environmental monitoring.

Microfluidics allows a fast and user-friendly approach for advanced monitoring due to its amenability for automation and miniaturization. Microfluidics is an outstandingly diverse and multidisciplinary field intersecting engineering, physics, chemistry, biotechnology, and nanotechnology. A hallmark of the advanced status of the technology is the integration of microfluidics and nucleic acid amplification tests, which offer promising point-of-care testing for accurate, rapid, and simple molecular analysis.

Point-of-care devices are leading to new possibilities in performing multiple and subsequent analyses in a single platform that includes sample preparation, amplification, and signal detection. The continuous effort to miniaturize integrated and automated sensing and diagnostic devices has promoted massive development in microfluidics. Although many challenges have arisen, we may achieve great success if we can overcome these differences, resulting in numerous current publications and commercial devices.

Any contributions that address the above are welcome to this Special Issue. Please feel free to contact me if you are interested in participating in this project or if you have any questions.

Dr. Kieu The Loan Trinh
Prof. Dr. Xianbo Lu
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Biosensors is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

microfluidics
biosensors
point-of-care
nucleic acid amplification tests
paper-based microfluidics
lab-on-a-chip
fabrication
sample preparation
Isothermal amplification
on-chip detection

Published Papers (6 papers)

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Editorial

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3 pages, 168 KiB

Open AccessEditorial

Microfluidic Biosensors for Point-of-Care Nucleic Acid Amplification Tests

by Kieu The Loan Trinh

Biosensors 2023, 13(1), 5; https://doi.org/10.3390/bios13010005 - 21 Dec 2022

Viewed by 1215

Abstract

This Special Issue of Biosensors, “Microfluidic Biosensors for Point-of-Care Nucleic Acid Amplification Tests” aims to gather original research papers and comprehensive reviews detailing novel research, fabrication methods, and applications, as well as the challenges and prospects of developing microfluidics for improved biosensing [...] Read more.

(This article belongs to the Special Issue Microfluidic Biosensors for Point-of-Care Nucleic Acid Amplification Tests)

Research

Jump to: Editorial

13 pages, 2361 KiB

Open AccessArticle

Oscillatory-Flow PCR Microfluidic Chip Driven by Low Speed Biaxial Centrifugation

by Yunlong Fan, Rongji Dai, Shuyu Lu, Xinyu Liu, Taiyan Zhou, Chunhua Yang, Xiaoming Hu, Xuefei Lv and Xiaoqiong Li

Biosensors 2023, 13(5), 555; https://doi.org/10.3390/bios13050555 - 18 May 2023

Cited by 2 | Viewed by 1388

Abstract

PCR is indispensable in basic science and biotechnology for in-orbit life science research. However, manpower and resources are limited in space. To address the constraints of in-orbit PCR, we proposed an oscillatory-flow PCR technique based on biaxial centrifugation. Oscillatory-flow PCR remarkably reduces the power requirements of the PCR process and has a relatively high ramp rate. A microfluidic chip that could perform dispensing, volume correction, and oscillatory-flow PCR of four samples simultaneously using biaxial centrifugation was designed. An automatic biaxial centrifugation device was designed and assembled to validate the biaxial centrifugation oscillatory-flow PCR. Simulation analysis and experimental tests indicated that the device could perform fully automated PCR amplification of four samples in one hour, with a ramp rate of 4.4

^{\circ}

C/s and average power consumption of less than 30 W. The PCR results were consistent with those obtained using conventional PCR equipment. Air bubbles generated during amplification were removed by oscillation. The chip and device realized a low-power, miniaturized, and fast PCR method under microgravity conditions, indicating good space application prospects and potential for higher throughput and extension to qPCR. Full article

(This article belongs to the Special Issue Microfluidic Biosensors for Point-of-Care Nucleic Acid Amplification Tests)

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12 pages, 3092 KiB

Open AccessArticle

Paper-Based Colorimetric Detection of miRNA-21 Using Pre-Activated Nylon Membrane and Peroxidase-Mimetic Activity of Cysteamine-Capped Gold Nanoparticles

by Maliana El Aamri, Hasna Mohammadi and Aziz Amine

Biosensors 2023, 13(1), 74; https://doi.org/10.3390/bios13010074 - 1 Jan 2023

Cited by 12 | Viewed by 2572

Abstract

Irregular expression of MicroRNA-21 (miRNA-21) is considered as a promising biomarker for early cancer diagnosis. In this paper, a new genosensor based on paper and nanozyme activity of cysteamine-capped gold nanoparticles (Cys/AuNPs) was developed to detect picomolar concentrations of miRNA-21. Such nanozyme catalyzes the colorimetric reaction of hydrogen peroxide (H₂O₂) and 3,3′,5,5′ tetramethylbenzidine (TMB), to produce a blue color measurable by a smartphone. Due to their positive charge, Cys/AuNPs were attached to the negative phosphate groups of the DNA strand backbone via electrostatic interactions, leading to the quantitative determination of miRNA-21 concentration by the peroxidase-like activity of Cys/AuNPs. Furthermore, a paper-based assay was carried out on nylon disk devices to allow fast immobilization of DNA_probe. After performing the paper-based assay, a good linear range was observed between 1 pM and 1 nM (Y = 0.080 [MiRNA-21]/pM + 13.846, R² = 0.993) with a detection limit of 0.5 pM. The developed method was effective, selective, and sensitive for the miRNA-21 detection. The application of the proposed method for miRNA-21 detection was examined in a human serum sample, and a recovery rate of 90.0–97.6% was obtained showing the acceptable accuracy of the developed approach. Full article

(This article belongs to the Special Issue Microfluidic Biosensors for Point-of-Care Nucleic Acid Amplification Tests)

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Graphical abstract

15 pages, 4730 KiB

Open AccessArticle

Comparison of Biosensing Methods Based on Different Isothermal Amplification Strategies: A Case Study with Erwinia amylovora

by Aleksandr V. Ivanov, Irina V. Safenkova, Natalia V. Drenova, Anatoly V. Zherdev and Boris B. Dzantiev

Biosensors 2022, 12(12), 1174; https://doi.org/10.3390/bios12121174 - 15 Dec 2022

Cited by 7 | Viewed by 2128

Abstract

Isothermal amplifications allow for the highly sensitive detection of nucleic acids, bypassing the use of instrumental thermal cycling. This work aimed to carry out an experimental comparison of the four most promising techniques: recombinase polymerase amplification (RPA) and loop-mediated isothermal amplification (LAMP) coupled with lateral flow test or coupled with additional amplification based on CRISPR/Cas12a resulting from the fluorescence of the Cas12a-cleaved probe. To compare the four amplification techniques, we chose the bacterial phytopathogen Erwinia amylovora (causative agent of fire blight), which has a quarantine significance in many countries and possesses a serious threat to agriculture. Three genes were chosen as the targets and primers were selected for each one (two for RPA and six for LAMP). They were functionalized by labels (biotin, fluorescein) at the 5′ ends for amplicons recognition by LFT. As a result, we developed LAMP-LFT, LAMP-CRISPR/Cas, RPA-LFT, and RPA-CRISPR/Cas for E. amylovora detection. The detection limit was 10⁴ CFU/mL for LAMP-LFT, 10³ CFU/mL for LAMP-CRISPR/Cas, and 10² CFU/mL for RPA-LFT and RPA-CRISPR/Cas. The results of four developed test systems were verified by qPCR on a panel of real samples. The developed assays based on RPA, LAMP, CRISPR/Cas12a, and LFT are rapid (30–55 min), user-friendly, and highly sensitive for E. amylovora detection. All proposed detection methods can be applied to fire blight diagnosis and effective management of this disease. Full article

(This article belongs to the Special Issue Microfluidic Biosensors for Point-of-Care Nucleic Acid Amplification Tests)

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11 pages, 774 KiB

Open AccessArticle

Rapid Detection of Attomolar SARS-CoV-2 Nucleic Acids in All-Dielectric Metasurface Biosensors

by Masanobu Iwanaga

Biosensors 2022, 12(11), 987; https://doi.org/10.3390/bios12110987 - 8 Nov 2022

Cited by 8 | Viewed by 1646

Abstract

Worldwide infection due to SARS-CoV-2 revealed that short-time and extremely high-sensitivity detection of nucleic acids is a crucial technique for human beings. Polymerase chain reactions have been mainly used for the SARS-CoV-2 detection over the years. However, an advancement in quantification of the detection and shortening runtime is important for present and future use. Here, we report a rapid detection scheme that is a combination of nucleic acid amplification and a highly efficient fluorescence biosensor, that is, a metasurface biosensor composed of a pair of an all-dielectric metasurface and a microfluidic transparent chip. In the present scheme, we show a series of proof-of-concept experimental results that the metasurface biosensors detected amplicons originating from attomolar SARS-CoV-2 nucleic acids and that the amplification was implemented within 1 h. Furthermore, this detection capability substantially satisfies an official requirement of 100 RNA copies/140 μL, which is a criterion for the reliable infection tests. Full article

(This article belongs to the Special Issue Microfluidic Biosensors for Point-of-Care Nucleic Acid Amplification Tests)

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10 pages, 1609 KiB

Open AccessArticle

Sensitive Electrochemical Biosensor for Rapid Screening of Tumor Biomarker TP53 Gene Mutation Hotspot

by Pengcheng Sun, Kai Niu, Haiying Du, Ruixin Li, Jiping Chen and Xianbo Lu

Biosensors 2022, 12(8), 658; https://doi.org/10.3390/bios12080658 - 19 Aug 2022

Cited by 4 | Viewed by 1946

Abstract

Rapid and sensitive detection of cancer biomarkers is crucial for cancer screening, early detection, and improving patient survival rate. The present study proposes an electrochemical gene-sensor capable of detecting tumor related TP53 gene mutation hotspots by self-assembly of sulfhydryl ended hairpin DNA probes tagged with methylene blue (MB) onto a gold electrode. By performing a hybridization reaction with the target DNA sequence, the gene-sensor can rearrange the probe’s structure, resulting in significant electrochemical signal differences by differential pulse voltammetry. When the DNA biosensor is hybridized with 1 μM target DNA, the peak current response signal can decrease more than 60%, displaying high sensitivity and specificity for the TP53 gene. The biosensor achieved rapid and sensitive detection of the TP53 gene with a detection limit of 10 nmol L⁻¹, and showed good specific recognition ability for single nucleotide polymorphism (SNP) and base sequence mismatches in the TP53 gene affecting residue 248 of the P53 protein. Moreover, the biosensor demonstrated good reproducibility, repeatability, operational stability, and anti-interference ability for target DNA molecule in the complex system of 50% fetal bovine serum. The proposed biosensor provides a powerful tool for the sensitive and specific detection of TP53 gene mutation hotspot sequences and could be used in clinical samples for early diagnosis and detection of cancer. Full article

(This article belongs to the Special Issue Microfluidic Biosensors for Point-of-Care Nucleic Acid Amplification Tests)

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