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Biosensors
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Advances in Amplification Methods for Biosensors
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Advances in Amplification Methods for Biosensors

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

Deadline for manuscript submissions: closed (31 January 2023) | Viewed by 25141

Special Issue Editor

Dr. Arnaud Buhot

E-Mail Website
Guest Editor
Grenoble Alpes University, CEA, CNRS, IRIG-SyMMES, 17 Rue des Martyrs, 38000 Grenoble, France
Interests: optoelectronic nose/tongue development; aptamer biosensors; surface plasmons resonance imaging; theory of microarrays (DNA or protein); biopolymer conformation; DNA based architectures; soft condensed matter
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Today, there is a rapidly growing demand for sensitive and selective biosensors in various domains, including environment monitoring such as (waste)water control, detection of pollution for personal/public safety, agricultural/food safety and quality control, veterinary and medical diagnostics, etc. For these applications, the main challenge remains to detect a minute amount of analytes in complex samples. Thus, recent biosensors based on the biomolecular recognition between analyte targets and relevant probes (antibodies, aptamers, molecular imprinted polymers (MIP), etc.) require the use of amplification methods to produce a measurable signal. The COVID-19 pandemic is a perfect demonstration with the need of (nucleic acids) amplification methods for the detection of SARS-CoV2 virus (RT-PCR, RT-LAMP, ELISA, etc.).

Therefore, this Special Issue of Biosensors will highlight recent advances in the design and development of novel amplification methods to improve the performance of biosensors. All the potential analyte targets will be considered ranging from molecules (ions, pesticides, hormones, antibiotics, endocrine disruptors, miRNA, proteins, biomarkers, etc.) to larger objects (viruses, spores, fungus, bacteria, cancer cells, etc.). Biosensors based on optical, electrochemical, chemiluminescence, fluorescence, resonant or mechanical transduction methods are encouraged. Finally, the amplification methods are not restricted: nucleic acid amplification (PCR, LAMP, RCA, NASBA, HCR, etc.) or logic gate circuits, enzymatic amplification (enzymes, DNAzymes, CRISPR, etc.), nanostructure-based amplification (nanoparticles, nanotubes, nanovesicles, MEMS/NEMS, nanosensors, etc.), as well as combined strategies implying several amplifications.

Dr. Arnaud Buhot
Guest Editor

If you want to learn more information or need any advice, you can contact the Special Issue Editor Jessica Zhou via <[email protected]> directly.

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

  • amplification
  • biosensors
  • electrochemical sensors
  • chemiluminescence sensors
  • optical sensors
  • resonant sensors
  • fluorescence sensors
  • nanosensors
  • nanoparticles
  • nanotubes
  • MEMS and/or NEMS
  • enzymes
  • DNAzymes
  • nucleic acid amplification
  • polymer chain reaction (PCR)
  • loop-mediated isothermal amplification (LAMP)
  • rolling circle amplification (RCA)
  • nucleic acid sequence based amplification (NASBA)
  • strand displacement amplification (SDA)
  • hybridization chain reaction (HCR)
  • recombinase polymerase amplification (RPA)
  • logic gate circuits
  • sensor analysis
  • environmental analysis
  • food quality control
  • diagnostics

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Published Papers (9 papers)

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Editorial

Jump to: Research, Review

3 pages, 171 KiB  
Editorial
Advances in Amplification Methods for Biosensors
by Arnaud Buhot
Biosensors 2023, 13(3), 365; https://doi.org/10.3390/bios13030365 - 10 Mar 2023
Viewed by 1477
Abstract
Today, there is a rapidly growing demand for sensitive and selective biosensors in various domains, including environmental monitoring such as (waste)water control, detection of pollution for personal/public safety, agricultural/food safety and quality control, veterinary and medical diagnostics, etc [...] Full article
(This article belongs to the Special Issue Advances in Amplification Methods for Biosensors)

Research

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12 pages, 3353 KiB  
Article
High-Performance FET-Based Dopamine-Sensitive Biosensor Platform Based on SOI Substrate
by Tae-Hwan Hyun and Won-Ju Cho
Biosensors 2023, 13(5), 516; https://doi.org/10.3390/bios13050516 - 3 May 2023
Cited by 4 | Viewed by 2592
Abstract
Dopamine is a catecholamine neurotransmitter that plays a significant role in the human central nervous system, even at extremely low concentrations. Several studies have focused on rapid and accurate detection of dopamine levels using field-effect transistor (FET)-based sensors. However, conventional approaches have poor [...] Read more.
Dopamine is a catecholamine neurotransmitter that plays a significant role in the human central nervous system, even at extremely low concentrations. Several studies have focused on rapid and accurate detection of dopamine levels using field-effect transistor (FET)-based sensors. However, conventional approaches have poor dopamine sensitivity with values <11 mV/log [DA]. Hence, it is necessary to increase the sensitivity of FET-based dopamine sensors. In the present study, we proposed a high-performance dopamine-sensitive biosensor platform based on dual-gate FET on a silicon-on-insulator substrate. This proposed biosensor overcame the limitations of conventional approaches. The biosensor platform consisted of a dual-gate FET transducer unit and a dopamine-sensitive extended gate sensing unit. The capacitive coupling between the top- and bottom-gate of the transducer unit allowed for self-amplification of the dopamine sensitivity, resulting in an increased sensitivity of 373.98 mV/log[DA] from concentrations 10 fM to 1 μM. Therefore, the proposed FET-based dopamine sensor is expected to be widely applied as a highly sensitive and reliable biosensor platform, enabling fast and accurate detection of dopamine levels in various applications such as medical diagnosis and drug development. Full article
(This article belongs to the Special Issue Advances in Amplification Methods for Biosensors)
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14 pages, 2871 KiB  
Article
Production of a Monoclonal Antibody for the Detection of Forchlorfenuron: Application in an Indirect Enzyme-Linked Immunosorbent Assay and Immunochromatographic Strip
by Xingmei Lei, A. M. Abd El-Aty, Lingyuan Xu, Jing Zhao, Jia Li, Song Gao, Yun Zhao, Yongxin She, Fen Jin, Jing Wang, Lufei Zheng, Maojun Jin and Bruce D. Hammock
Biosensors 2023, 13(2), 239; https://doi.org/10.3390/bios13020239 - 7 Feb 2023
Cited by 7 | Viewed by 2286
Abstract
In this study, a monoclonal antibody (mAb) specific to forchlorfenuron (CPPU) with high sensitivity and specificity was produced and designated (9G9). To detect CPPU in cucumber samples, an indirect enzyme-linked immunosorbent assay (ic-ELISA) and a colloidal gold nanobead immunochromatographic test strip (CGN-ICTS) were [...] Read more.
In this study, a monoclonal antibody (mAb) specific to forchlorfenuron (CPPU) with high sensitivity and specificity was produced and designated (9G9). To detect CPPU in cucumber samples, an indirect enzyme-linked immunosorbent assay (ic-ELISA) and a colloidal gold nanobead immunochromatographic test strip (CGN-ICTS) were established using 9G9. The half-maximal inhibitory concentration (IC50) and the LOD for the developed ic-ELISA were determined to be 0.19 ng/mL and 0.04 ng/mL in the sample dilution buffer, respectively. The results indicate that the sensitivity of the antibodies prepared in this study (9G9 mAb) was higher than those reported in the previous literature. On the other hand, in order to achieve rapid and accurate detection of CPPU, CGN-ICTS is indispensable. The IC50 and the LOD for the CGN-ICTS were determined to be 27 ng/mL and 6.1 ng/mL. The average recoveries of the CGN-ICTS ranged from 68 to 82%. The CGN-ICTS and ic-ELISA quantitative results were all confirmed by liquid chromatography—tandem mass spectrometry (LC-MS/MS) with 84–92% recoveries, which indicated the methods developed herein are appropriate for detecting CPPU in cucumber. The CGN-ICTS method is capable of both qualitative and semiquantitative analysis of CPPU, which makes it a suitable alternative complex instrument method for on-site detection of CPPU in cucumber samples since it does not require specialized equipment. Full article
(This article belongs to the Special Issue Advances in Amplification Methods for Biosensors)
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14 pages, 2690 KiB  
Article
Exploiting the Nucleic Acid Nature of Aptamers for Signal Amplification
by Miriam Jauset-Rubio, Mayreli Ortiz and Ciara K. O’Sullivan
Biosensors 2022, 12(11), 972; https://doi.org/10.3390/bios12110972 - 4 Nov 2022
Cited by 4 | Viewed by 2305
Abstract
Aptamer-based assays and sensors are garnering increasing interest as alternatives to antibodies, particularly due to their increased flexibility for implementation in alternative assay formats, as they can be employed in assays designed for nucleic acids, such as molecular aptamer beacons or aptamer detection [...] Read more.
Aptamer-based assays and sensors are garnering increasing interest as alternatives to antibodies, particularly due to their increased flexibility for implementation in alternative assay formats, as they can be employed in assays designed for nucleic acids, such as molecular aptamer beacons or aptamer detection combined with amplification. In this work, we took advantage of the inherent nucleic acid nature of aptamers to enhance sensitivity in a rapid and facile assay format. An aptamer selected against the anaphylactic allergen β-conglutin was used to demonstrate the proof of concept. The aptamer was generated by using biotinylated dUTPs, and the affinity of the modified aptamer as compared to the unmodified aptamer was determined by using surface plasmon resonance to calculate the dissociation constant (KD), and no significant improvement in affinity due to the incorporation of the hydrophobic biotin was observed. The modified aptamer was then applied in a colorimetric competitive enzyme-linked oligonucleotide assay, where β-conglutin was immobilized on the wells of a microtiter plate, competing with β-conglutin free in solution for the binding to the aptamer. The limit of detection achieved was 68 pM, demonstrating an improvement in detection limit of three orders of magnitude as compared with the aptamer simply modified with a terminal biotin label. The concept was then exploited by using electrochemical detection and screen-printed electrodes where detection limits of 326 fM and 7.89 fM were obtained with carbon and gold electrodes, respectively. The assay format is generic in nature and can be applied to all aptamers, facilitating an easy and cost-effective means to achieve lower detection limits. Full article
(This article belongs to the Special Issue Advances in Amplification Methods for Biosensors)
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13 pages, 1619 KiB  
Article
Efficient Entrapment of Alpha-Synuclein Biotinylated Antibody in KCC-1-NH-CS2 and Application for the Sensitive Diagnosis of Parkinson’s Using Recognition of Biomarker: An Innovative Electrochemical Label-Free Immunosensor for the Biomedical Analysis of Neurodegenerative Diseases
by Hossein Navay Baghban, Mohammad Hasanzadeh, Yuqian Liu and Farzad Seidi
Biosensors 2022, 12(10), 911; https://doi.org/10.3390/bios12100911 - 21 Oct 2022
Cited by 6 | Viewed by 2349
Abstract
The early detection of Parkinson’s disease (PD) is a critical issue in terms of efficiency. Alpha-synuclein (α-Syn) is a biomarker in PD checks. Alpha-synuclein (α-syn) is the major constituent of Lewy bodies and a pathogenic hallmark of all synucleinopathies, including PDs, dementia with [...] Read more.
The early detection of Parkinson’s disease (PD) is a critical issue in terms of efficiency. Alpha-synuclein (α-Syn) is a biomarker in PD checks. Alpha-synuclein (α-syn) is the major constituent of Lewy bodies and a pathogenic hallmark of all synucleinopathies, including PDs, dementia with Lewy bodies, and multiple system atrophy. In this study, KCC-1-NH-CS2 was conjugated with biotinylated Ab and entrapped in P(β-CD) polymer cavities. Using this approach, a novel electrochemical label-free immunosensor was designed for the quantification of α-syn in real human samples. For this purpose, the glassy carbon electrode electropolymerized with P(β-CD) biopolymer provided an excellent matrix for entrapping of KCC-1-NH-CS2 loaded with the biotinylated antibody of α-syn. Using the chronoamperometric technique, the proposed immunosensor shows a suitable range of 0.02 to 64 ng/mL for the determination of α-syn. Additionally, a low limit of quantification of the engineered biosensor was obtained at 0.02 ng/mL. The developed immunosensor’s adequate stability, sensitivity, and selectivity, together with its ease of manufacture, make it a promising diagnostic technique for further research. This study also will pave the way for further applications of the synergetic effect of β-CD and KCC-1-NH-CS2 for biomedical analysis in the near future. Full article
(This article belongs to the Special Issue Advances in Amplification Methods for Biosensors)
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14 pages, 4967 KiB  
Article
An Innovative Sandwich Type Biosensor towards Sensitive and Selective Monitoring of 2-Arachidonoylglycerol in Human Plasma Samples Using P(β-CD)-AuNPs-DDT as Amplificant Agent: A New Immuno-Platform for the Recognition of Endocannabinoids in Real Samples
by Nastaran Aletaha, Kambiz Ghaseminasab, Mohammad Hasanzadeh, Fereshteh Kohansal, Yuqian Liu and Farzad Seidi
Biosensors 2022, 12(10), 791; https://doi.org/10.3390/bios12100791 - 26 Sep 2022
Cited by 4 | Viewed by 2241
Abstract
In this work, 2-AG was successfully detected in human plasma samples using a new sandwich-type electrochemical immune device based on poly-β-cyclodextrin P(β-CD) functionalized with AuNPs-DDT and toluidine blue. The P(β-CD) ensured the bioactivity and stability of the [...] Read more.
In this work, 2-AG was successfully detected in human plasma samples using a new sandwich-type electrochemical immune device based on poly-β-cyclodextrin P(β-CD) functionalized with AuNPs-DDT and toluidine blue. The P(β-CD) ensured the bioactivity and stability of the immobilized 2-AG antibody by providing a broad surface for the efficient immobilization of the biotinylated antibody. To complete the top section of the immunosensor (reporter), an HRP-conjugated antibody of 2-AG (secondary antibody (Ab2)) was attached to the surface of a glassy carbon electrode (GCE) modified by P(β-CD), as well as a primarily biotinylated antibody (Ab1). The biosensor fabrication process was monitored using field-emission scanning electron microscope (FE-SEM) and EDS methods. Using the differential pulse voltammetry technique, the immunosensor was utilized for detection of 2-AG in real samples. The suggested interface increased the surface area, which allowed for the immobilization of a large quantity of anti-2-AG antibody while also improving biocompatibility, stability, and electrical conductivity. Finally, the suggested immunosensor’s limit of quantitation was determined to be 0.0078 ng/L, with a linear range of 0.0078 to 1.0 ng/L. The results showed that the suggested bioassay can be utilized for diagnosis of 2-AG in clinical samples as a unique and ultrasensitive electrochemical biodevice. Full article
(This article belongs to the Special Issue Advances in Amplification Methods for Biosensors)
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12 pages, 2737 KiB  
Article
Hundreds-Dollar-Level Multiplex Integrated RT-qPCR Quantitative System for Field Detection
by Zhihao Lan, Yu Guo, Kangning Wang, Yipeng Zhang, Youyun Chen, Dezhou Zheng, Xiaolong Xu and Wenming Wu
Biosensors 2022, 12(9), 706; https://doi.org/10.3390/bios12090706 - 1 Sep 2022
Cited by 2 | Viewed by 2616
Abstract
The COVID-19 pandemic poses a threat to global health. Due to its high sensitivity, specificity, and stability, real-time fluorescence quantitative (real-time PCR) detection has become the most extensively used approach for diagnosing SARS-CoV-2 pneumonia. According to a report from the World Health Organization, [...] Read more.
The COVID-19 pandemic poses a threat to global health. Due to its high sensitivity, specificity, and stability, real-time fluorescence quantitative (real-time PCR) detection has become the most extensively used approach for diagnosing SARS-CoV-2 pneumonia. According to a report from the World Health Organization, emerging and underdeveloped nations lack nucleic acid detection kits and polymerase chain reaction (PCR) instruments for molecular biological detection. In addition, sending samples to a laboratory for testing may result in considerable delays between sampling and diagnosis, which is not favorable to the timely prevention and control of new crown outbreaks. Concurrently, there is an urgent demand for accurate PCR devices that do not require a laboratory setting, are more portable, and are capable of completing testing on-site. Hence, we report on HDLRT-qPCR, a new, low-cost, multiplexed real-time fluorescence detection apparatus that we have developed for on-site testing investigations of diverse diseases in developing nations. This apparatus can complete on-site testing rapidly and sensitively. The entire cost of this instrument does not exceed USD 760. In order to demonstrate the applicability of our PCR instrument, we conducted testing that revealed that we achieved gradient amplification and melting curves comparable to those of commercially available equipment. Good consistency characterized the testing outcomes. The successful detection of target genes demonstrates the reliability of our inexpensive PCR diagnostic technique. With this apparatus, there is no need to transport samples to a central laboratory; instead, we conduct testing at the sampling site. This saves time on transportation, substantially accelerates overall testing speed, and provides results within 40 min. Full article
(This article belongs to the Special Issue Advances in Amplification Methods for Biosensors)
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18 pages, 5287 KiB  
Article
Kinetics of Isothermal Dumbbell Exponential Amplification: Effects of Mix Composition on LAMP and Its Derivatives
by Maud Savonnet, Mathilde Aubret, Patricia Laurent, Yoann Roupioz, Myriam Cubizolles and Arnaud Buhot
Biosensors 2022, 12(5), 346; https://doi.org/10.3390/bios12050346 - 18 May 2022
Cited by 8 | Viewed by 3451
Abstract
Loop-mediated isothermal amplification (LAMP) is an exponential amplification method of DNA strands that is more and more used for its high performances. Thanks to its high sensitivity and selectivity, LAMP found numerous applications from the detection of pathogens or viruses through their genome [...] Read more.
Loop-mediated isothermal amplification (LAMP) is an exponential amplification method of DNA strands that is more and more used for its high performances. Thanks to its high sensitivity and selectivity, LAMP found numerous applications from the detection of pathogens or viruses through their genome amplification to its incorporation as an amplification strategy in protein or miRNA biomarker quantification. The LAMP method is composed of two stages: the first one consists in the transformation of the DNA strands into dumbbell structures formed of two stems and loops thanks to four primers; then, in the second stage, only two primers are required to amplify the dumbbells exponentially in numerous hairpins of increasing lengths. In this paper, we propose a theoretical framework to analyze the kinetics of the second stage of LAMP, the isothermal dumbbell exponential amplification (IDEA) as function of the physico-chemical parameters of the amplification reaction. Dedicated experiments validate the models. We believe these results may help the optimization of LAMP performances by reducing the number of experiments necessary to find the best parameters. Full article
(This article belongs to the Special Issue Advances in Amplification Methods for Biosensors)
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Review

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31 pages, 4043 KiB  
Review
Emerging Multiplex Nucleic Acid Diagnostic Tests for Combating COVID-19
by Patarajarin Akarapipad, Elizabeth Bertelson, Alexander Pessell, Tza-Huei Wang and Kuangwen Hsieh
Biosensors 2022, 12(11), 978; https://doi.org/10.3390/bios12110978 - 7 Nov 2022
Cited by 9 | Viewed by 4223
Abstract
The COVID-19 pandemic caused by SARS-CoV-2 has drawn attention to the need for fast and accurate diagnostic testing. Concerns from emerging SARS-CoV-2 variants and other circulating respiratory viral pathogens further underscore the importance of expanding diagnostic testing to multiplex detection, as single-plex diagnostic [...] Read more.
The COVID-19 pandemic caused by SARS-CoV-2 has drawn attention to the need for fast and accurate diagnostic testing. Concerns from emerging SARS-CoV-2 variants and other circulating respiratory viral pathogens further underscore the importance of expanding diagnostic testing to multiplex detection, as single-plex diagnostic testing may fail to detect emerging variants and other viruses, while sequencing can be too slow and too expensive as a diagnostic tool. As a result, there have been significant advances in multiplex nucleic-acid-based virus diagnostic testing, creating a need for a timely review. This review first introduces frequent nucleic acid targets for multiplex virus diagnostic tests, then proceeds to a comprehensive and up-to-date overview of multiplex assays that incorporate various detection reactions and readout modalities. The performances, advantages, and disadvantages of these assays are discussed, followed by highlights of platforms that are amenable for point-of-care use. Finally, this review points out the remaining technical challenges and shares perspectives on future research and development. By examining the state of the art and synthesizing existing development in multiplex nucleic acid diagnostic tests, this review can provide a useful resource for facilitating future research and ultimately combating COVID-19. Full article
(This article belongs to the Special Issue Advances in Amplification Methods for Biosensors)
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