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Biosensors, Volume 10, Issue 5 (May 2020) – 11 articles

Cover Story (view full-size image): The sensitivity of aptamer-based assays is ultimately limited by the functional affinity of the synthetic receptors. In this work, we show how the use of isothermal nucleic acid amplification increases the sensitivity and the apparent affinity of aptamers, even when weak aptamers are employed. Specifically, we compare terminal deoxynucleotidyl transferase (TdT) and rolling circle amplification (RCA) in combination with two different aptamers against alpha-fetoprotein. Both DNA amplifications are useful although the improvement depends on the true affinity of the binding pair, which ultimately limits the analytical usefulness.View this paper.
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18 pages, 2581 KiB  
Review
Recent Progress on the Electrochemical Biosensing of Escherichia coli O157:H7: Material and Methods Overview
by Nasrin Razmi, Mohammad Hasanzadeh, Magnus Willander and Omer Nur
Biosensors 2020, 10(5), 54; https://doi.org/10.3390/bios10050054 - 18 May 2020
Cited by 31 | Viewed by 7275
Abstract
Escherichia coli O157:H7 (E. coli O157:H7) is a pathogenic strain of Escherichia coli which has issued as a public health threat because of fatal contamination of food and water. Therefore, accurate detection of pathogenic E. coli is important in environmental and food [...] Read more.
Escherichia coli O157:H7 (E. coli O157:H7) is a pathogenic strain of Escherichia coli which has issued as a public health threat because of fatal contamination of food and water. Therefore, accurate detection of pathogenic E. coli is important in environmental and food quality monitoring. In spite of their advantages and high acceptance, culture-based methods, enzyme-linked immunosorbent assays (ELISAs), polymerase chain reaction (PCR), flow cytometry, ATP bioluminescence, and solid-phase cytometry have various drawbacks, including being time-consuming, requiring trained technicians and/or specific equipment, and producing biological waste. Therefore, there is necessity for affordable, rapid, and simple approaches. Electrochemical biosensors have shown great promise for rapid food- and water-borne pathogen detection. Over the last decade, various attempts have been made to develop techniques for the rapid quantification of E. coli O157:H7. This review covers the importance of E. coli O157:H7 and recent progress (from 2015 to 2020) in the development of the sensitivity and selectivity of electrochemical sensors developed for E. coli O157:H7 using different nanomaterials, labels, and electrochemical transducers. Full article
(This article belongs to the Special Issue Electrochemical Biosensors)
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13 pages, 3683 KiB  
Article
Silver SERS Adenine Sensors with a Very Low Detection Limit
by Yonhua Tzeng and Bo-Yi Lin
Biosensors 2020, 10(5), 53; https://doi.org/10.3390/bios10050053 - 15 May 2020
Cited by 22 | Viewed by 4490
Abstract
The detection of adenine molecules at very low concentrations is important for biological and medical research and applications. This paper reports a silver-based surface-enhanced Raman scattering (SERS) sensor with a very low detection limit for adenine molecules. Clusters of closely packed silver nanoparticles [...] Read more.
The detection of adenine molecules at very low concentrations is important for biological and medical research and applications. This paper reports a silver-based surface-enhanced Raman scattering (SERS) sensor with a very low detection limit for adenine molecules. Clusters of closely packed silver nanoparticles on surfaces of discrete ball-like copper bumps partially covered with graphene are deposited by immersion in silver nitrate. These clusters of silver nanoparticles exhibit abundant nanogaps between nanoparticles, where plasmonic coupling induces very high local electromagnetic fields. Silver nanoparticles growing perpendicularly on ball-like copper bumps exhibit surfaces of large curvature, where electromagnetic field enhancement is high. Between discrete ball-like copper bumps, the local electromagnetic field is low. Silver is not deposited on the low-field surface area. Adenine molecules interact with silver by both electrostatic and functional groups and exhibit low surface diffusivity on silver surface. Adenine molecules are less likely to adsorb on low-field sensor surface without silver. Therefore, adenine molecules have a high probability of adsorbing on silver surface of high local electric fields and contribute to the measured Raman scattering signal strength. We demonstrated SERS sensors made of clusters of silver nanoparticles deposited on discrete ball-like copper bumps with very a low detection limit for detecting adenine water solution of a concentration as low as 10−11 M. Full article
(This article belongs to the Special Issue SERS-Spectroscopy for Biosensing)
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14 pages, 1197 KiB  
Article
Paper-Based Working Electrodes Coated with Mercury or Bismuth Films for Heavy Metals Determination
by Alberto Sánchez-Calvo, Maria Carmen Blanco-López and Agustín Costa-García
Biosensors 2020, 10(5), 52; https://doi.org/10.3390/bios10050052 - 13 May 2020
Cited by 25 | Viewed by 4534
Abstract
Paper-based carbon working electrodes were modified with mercury or bismuth films for the determination of trace metals in aqueous solutions. Both modification procedures were optimized in terms of selectivity and sensitivity for the determination of different heavy metals, aiming their simultaneous determination. Cd [...] Read more.
Paper-based carbon working electrodes were modified with mercury or bismuth films for the determination of trace metals in aqueous solutions. Both modification procedures were optimized in terms of selectivity and sensitivity for the determination of different heavy metals, aiming their simultaneous determination. Cd (II), Pb (II) and In (III) could be quantified with both films. However, Cu (II) could not be determined with bismuth films. The modification with mercury films led to the most sensitive method, with linear ranges between 0.1 and 10 µg/mL and limits of detection of 0.4, 0.1, 0.04 and 0.2 µg/mL for Cd (II), Pb (II), In (III) and Cu (II), respectively. Nevertheless, the bismuth film was a more sustainable alternative to mercury. Tap-water samples were analyzed for the determination of metals by standard addition methodology with good accuracy, by using a low-cost and easily disposable paper-based electrochemical platform. This system demonstrated its usefulness for monitoring heavy metals in water. Full article
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19 pages, 1358 KiB  
Review
Yeast-Based Biosensors: Current Applications and New Developments
by Helene Martin-Yken
Biosensors 2020, 10(5), 51; https://doi.org/10.3390/bios10050051 - 13 May 2020
Cited by 26 | Viewed by 7266
Abstract
Biosensors are regarded as a powerful tool to detect and monitor environmental contaminants, toxins, and, more generally, organic or chemical markers of potential threats to human health. They are basically composed of a sensor part made up of either live cells or biological [...] Read more.
Biosensors are regarded as a powerful tool to detect and monitor environmental contaminants, toxins, and, more generally, organic or chemical markers of potential threats to human health. They are basically composed of a sensor part made up of either live cells or biological active molecules coupled to a transducer/reporter technological element. Whole-cells biosensors may be based on animal tissues, bacteria, or eukaryotic microorganisms such as yeasts and microalgae. Although very resistant to adverse environmental conditions, yeasts can sense and respond to a wide variety of stimuli. As eukaryotes, they also constitute excellent cellular models to detect chemicals and organic contaminants that are harmful to animals. For these reasons, combined with their ease of culture and genetic modification, yeasts have been commonly used as biological elements of biosensors since the 1970s. This review aims first at giving a survey on the different types of yeast-based biosensors developed for the environmental and medical domains. We then present the technological developments currently undertaken by academic and corporate scientists to further drive yeasts biosensors into a new era where the biological element is optimized in a tailor-made fashion by in silico design and where the output signals can be recorded or followed on a smartphone. Full article
(This article belongs to the Section Biosensor and Bioelectronic Devices)
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14 pages, 999 KiB  
Article
Fecal Volatile Organic Compound Profiles are Not Influenced by Gestational Age and Mode of Delivery: A Longitudinal Multicenter Cohort Study
by Nancy Deianova, Sofia el Manouni el Hassani, Hendrik J. Niemarkt, Veerle Cossey, Anton H. van Kaam, Floor Jenken, Mirjam M. van Weissenbruch, Esmee M. Doedes, Kyra Baelde, Renee Menezes, Marc A. Benninga, Wouter J. de Jonge, Nanne K. de Boer and Tim G. de Meij
Biosensors 2020, 10(5), 50; https://doi.org/10.3390/bios10050050 - 11 May 2020
Cited by 8 | Viewed by 4144
Abstract
Fecal volatile organic compounds (VOC) reflect human and gut microbiota metabolic pathways and their interaction. VOC behold potential as non-invasive preclinical diagnostic biomarkers in various diseases, e.g., necrotizing enterocolitis and late onset sepsis. There is a need for standardization and assessment of the [...] Read more.
Fecal volatile organic compounds (VOC) reflect human and gut microbiota metabolic pathways and their interaction. VOC behold potential as non-invasive preclinical diagnostic biomarkers in various diseases, e.g., necrotizing enterocolitis and late onset sepsis. There is a need for standardization and assessment of the influence of clinical and environmental factors on the VOC outcome before this technique can be applied in clinical practice. The aim of this study was to investigate the influence of gestational age (GA) and mode of delivery on the fecal VOC pattern in preterm infants born below 30 weeks of gestation. Longitudinal fecal samples, collected on days 7, 14, and 21 postnatally, were analyzed by an electronic nose device (Cyranose 320®). In total, 58 preterm infants were included (29 infants born at GA 24–26 weeks vs. 29 at 27–29 completed weeks, 24 vaginally born vs. 34 via C-section). No differences were identified at any predefined time point in terms of GA and delivery mode (p > 0.05). We, therefore, concluded that correction for these factors in this population is not warranted when performing fecal VOC analysis in the first three weeks of life. Full article
(This article belongs to the Special Issue Noninvasive Early Disease Diagnosis)
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11 pages, 3025 KiB  
Article
Battery-Powered Portable Rotary Real-Time Fluorescent qPCR with Low Energy Consumption, Low Cost, and High Throughput
by Limin He, Benliang Sang and Wenming Wu
Biosensors 2020, 10(5), 49; https://doi.org/10.3390/bios10050049 - 08 May 2020
Cited by 14 | Viewed by 4616
Abstract
The traditional qPCR instrument is bulky, expensive, and inconvenient to carry, so we report a portable rotary real-time fluorescent PCR (polymerase chain reaction) that completes the PCR amplification of DNA in the field, and the reaction can be observed in real-time. Through the [...] Read more.
The traditional qPCR instrument is bulky, expensive, and inconvenient to carry, so we report a portable rotary real-time fluorescent PCR (polymerase chain reaction) that completes the PCR amplification of DNA in the field, and the reaction can be observed in real-time. Through the analysis of a target gene, namely pGEM-3Zf (+), the gradient amplification and melting curves are compared to commercial devices. The results confirm the stability of our device. This is the first use of a mechanical rotary structure to achieve gradient amplification curves and melting curves comparable to commercial instruments. The average power consumption of our system is about 7.6 W, which is the lowest energy consumption for real-time fluorescence quantification in shunting PCR and enables the use of our device in the field thanks to its self-contained power supply based on a lithium battery. In addition, all of the equipment costs only about 710 dollars, which is far lower than the cost of a commercial PCR instrument because the control system through mechanical displacement replaces the traditional TEC (thermoelectric cooler) temperature control. Moreover, the equipment has a low technical barrier, which can suit the needs of non-professional settings, with strong repeatability. Full article
(This article belongs to the Special Issue Fluorescence Biosensors 2020)
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9 pages, 446 KiB  
Article
Smell—Adding a New Dimension to Urinalysis
by Eva H. Visser, Daan J. C. Berkhout, Jiwanjot Singh, Annemieke Vermeulen, Niloufar Ashtiani, Nanne K. de Boer, Joanna A. E. van Wijk, Tim G. de Meij and Arend Bökenkamp
Biosensors 2020, 10(5), 48; https://doi.org/10.3390/bios10050048 - 05 May 2020
Cited by 6 | Viewed by 4010
Abstract
Background: Urinary tract infections (UTI) are among the most common infections in children. The primary tool to detect UTI is dipstick urinalysis; however, this has limited sensitivity and specificity. Therefore, urine culture has to be performed to confirm a UTI. Urinary volatile organic [...] Read more.
Background: Urinary tract infections (UTI) are among the most common infections in children. The primary tool to detect UTI is dipstick urinalysis; however, this has limited sensitivity and specificity. Therefore, urine culture has to be performed to confirm a UTI. Urinary volatile organic compounds (VOC) may serve as potential biomarker for diagnosing UTI. Previous studies on urinary VOCs focused on detection of UTI in a general population; therefore, this proof-of-principle study was set up in a clinical high-risk pediatric population. Methods: This study was performed at a tertiary nephro-urological clinic. Patients included were 0–18 years, clinically suspected of a UTI, and had abnormal urinalysis. Urine samples were divided into four groups, i.e., urine without bacterial growth, contamination, colonization, and UTI. VOC analysis was performed using an electronic nose (eNose) (Cyranose 320®) and VOC profiles of subgroups were compared. Results: Urinary VOC analysis discriminated between UTI and non-UTI samples (AUC 0.70; p = 0.048; sensitivity 0.67, specificity 0.70). The diagnostic accuracy of VOCs improved when comparing urine without bacterial growth versus with UTI (AUC 0.80; p = 0.009, sensitivity 0.79, specificity 0.75). Conclusions: In an intention-to-diagnose high-risk pediatric population, UTI could be discriminated from non-UTI by VOC profiling, using an eNose. Since eNose can be used as bed-side test, these results suggest that urinary VOC analysis may serve as an adjuvant in the diagnostic work-up of UTI in children. Full article
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10 pages, 1633 KiB  
Article
An Array of MOX Sensors and ANNs to Assess Grated Parmigiano Reggiano Cheese Packs’ Compliance with CFPR Guidelines
by Marco Abbatangelo, Estefanía Núñez-Carmona, Veronica Sberveglieri, Dario Zappa, Elisabetta Comini and Giorgio Sberveglieri
Biosensors 2020, 10(5), 47; https://doi.org/10.3390/bios10050047 - 02 May 2020
Cited by 7 | Viewed by 3650
Abstract
Parmigiano Reggiano cheese is one of the most appreciated Italian foods on account of its high nutrient content and taste. Due to its high cost, these characteristics make this product subject to counterfeiting in different forms. In this study, an approach based on [...] Read more.
Parmigiano Reggiano cheese is one of the most appreciated Italian foods on account of its high nutrient content and taste. Due to its high cost, these characteristics make this product subject to counterfeiting in different forms. In this study, an approach based on an array of gas sensors has been employed to assess if it was possible to distinguish different samples based on their aroma. Samples were characterized in terms of rind percentage, seasoning, and rind working process. From the responses of the sensors, five features were extracted and the capability of these parameters to recognize target classes was tested with statistical analysis. Hence, the performance of the sensors’ array was quantified using artificial neural networks. To simplify the problem, a hierarchical approach has been used: three steps of classification were performed, and in each step one parameter of the grated cheese was identified (firstly, seasoning; secondly, rind working process; finally, rind percentage). The accuracies ranged from 88.24% to 100%. Full article
(This article belongs to the Special Issue Sensors for Food Safety)
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11 pages, 2408 KiB  
Article
On the Electrochemical Detection of Alpha-Fetoprotein Using Aptamers: DNA Isothermal Amplification Strategies to Improve the Performance of Weak Aptamers
by Ramón Lorenzo-Gómez, Daniel González-Robles, Rebeca Miranda-Castro, Noemí de-los-Santos-Álvarez and María Jesús Lobo-Castañón
Biosensors 2020, 10(5), 46; https://doi.org/10.3390/bios10050046 - 30 Apr 2020
Cited by 6 | Viewed by 5009
Abstract
Affinity characterization is essential to develop reliable aptamers for tumor biomarker detection. For alpha-fetoprotein (AFP), a biomarker of hepatocellular carcinoma (HCC), two DNA aptamers were described with very different affinity. In this work, we estimate the dissociation constant of both of them by [...] Read more.
Affinity characterization is essential to develop reliable aptamers for tumor biomarker detection. For alpha-fetoprotein (AFP), a biomarker of hepatocellular carcinoma (HCC), two DNA aptamers were described with very different affinity. In this work, we estimate the dissociation constant of both of them by means of a direct assay on magnetic beads modified with AFP and electrochemical detection on carbon screen-printed electrodes (SPCE). Unlike previous works, both aptamers showed similar dissociation constant (Kd) values, in the subµM range. In order to improve the performance of these aptamers, we proposed the isothermal amplification of the aptamers by both terminal deoxynucleotidyl transferase (TdT) and rolling circle amplification (RCA). Both DNA amplifications improved the sensitivity and also the apparent binding constants from 713 nM to 189 nM for the short aptamer and from 526 nM to 32 nM for the long aptamer. This improvement depends on the true affinity of the binding pair, which ultimately limits the analytical usefulness. Full article
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50 pages, 10450 KiB  
Review
Immobilization Techniques for Aptamers on Gold Electrodes for the Electrochemical Detection of Proteins: A Review
by Franziska V. Oberhaus, Dieter Frense and Dieter Beckmann
Biosensors 2020, 10(5), 45; https://doi.org/10.3390/bios10050045 - 28 Apr 2020
Cited by 76 | Viewed by 13643
Abstract
The development of reliable biosensing platforms plays a key role in the detection of proteins in clinically and environmentally derived samples for diagnostics, as well as for process monitoring in biotechnological productions. For this purpose, the biosensor has to be stable and reproducible, [...] Read more.
The development of reliable biosensing platforms plays a key role in the detection of proteins in clinically and environmentally derived samples for diagnostics, as well as for process monitoring in biotechnological productions. For this purpose, the biosensor has to be stable and reproducible, and highly sensitive to detect potentially extremely low concentrations and prevent the nonspecific binding of interfering compounds. In this review, we present an overview of recently published (2017–2019) immobilization techniques for aptamers on gold electrodes for the electrochemical detection of proteins. These include the direct immobilization of thiolated aptamers and the utilization of short linkers, streptavidin/biotin interaction, as well as DNA nanostructures and reduced graphene oxide as immobilization platforms. Applied strategies for signal amplification and the prevention of biofouling are additionally discussed, as they play a crucial role in the design of biosensors. While a wide variety of amplification strategies are already available, future investigations should aim to establish suitable antifouling strategies that are compatible with electrochemical measurements. The focus of our review lies on the detailed discussion of the underlying principles and the presentation of utilized chemical protocols in order to provide the reader with promising ideas and profound knowledge of the subject, as well as an update on recent discoveries and achievements. Full article
(This article belongs to the Section Biosensor and Bioelectronic Devices)
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7 pages, 2183 KiB  
Communication
Host-Modulation Therapy and Chair-Side Diagnostics in the Treatment of Peri-Implantitis
by Timo Sorsa, Joseph Bacigalupo, Mauno Könönen, Pirjo Pärnänen and Ismo T. Räisänen
Biosensors 2020, 10(5), 44; https://doi.org/10.3390/bios10050044 - 25 Apr 2020
Cited by 19 | Viewed by 4699
Abstract
Previous studies report periodontitis and peri-implantitis being able to induce systemic low-grade inflammation, which is known to be associated with increased risk for some systemic medical disease such as cardiovascular disease. In this regard, recent studies have shown that host modulation therapy (HMT) [...] Read more.
Previous studies report periodontitis and peri-implantitis being able to induce systemic low-grade inflammation, which is known to be associated with increased risk for some systemic medical disease such as cardiovascular disease. In this regard, recent studies have shown that host modulation therapy (HMT) together with traditional mechanical and surgical treatment not only cease the progression of periodontitis but also reduce the systemic collagenolytic biomarkers in both oral fluids and circulation. This suggests that the corresponding adjunctive HMT-medication could be effective in the prevention and treatment of dental peri-implantitis, as well. Furthermore, low-cost, safe, and practical oral fluid active matrix metalloproteinase-8 (aMMP-8) lateral-flow immunotests have been proposed as point-of-care/chair-side diagnostic tools to detect peri-implantitis and periodontitis, and to monitor their effective resolutions, while using various therapeutic strategies, including host modulation. This study reports the potential benefits of HMT-medication in the prevention and treatment of dental peri-implantitis among five patients (four of five were current/ex-smokers). In addition, the aMMP-8 point-of-care test diagnosed 20 peri-implantitis and 20 healthy controls correctly. In conclusion, this study and previous studies support the potential effectiveness of HMT-medication(s) and point-of-care/chair-side technologies in the treatment and diagnostics/monitoring of peri-implantitis. However, more studies are needed to further confirm this. Full article
(This article belongs to the Section Biosensors and Healthcare)
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