Research

Jump to: Review

14 pages, 2987 KiB

Open AccessArticle

Research on a Magnetic Separation-Based Rapid Nucleic Acid Extraction System and Its Detection Applications

by Yao Li, Sha Liu, Yuanyuan Wang, Yue Wang, Song Li, Nongyue He, Yan Deng and Zhu Chen

Biosensors 2023, 13(10), 903; https://doi.org/10.3390/bios13100903 - 23 Sep 2023

Cited by 1 | Viewed by 1555

Nucleic acid extraction represents the “first step” in molecular diagnostic experiments. The quality of this extraction serves as a fundamental prerequisite for ensuring the accuracy of nucleic acid detection. This article presents a comprehensive design scheme for a rapid automated nucleic acid extraction [...] Read more.

Nucleic acid extraction represents the “first step” in molecular diagnostic experiments. The quality of this extraction serves as a fundamental prerequisite for ensuring the accuracy of nucleic acid detection. This article presents a comprehensive design scheme for a rapid automated nucleic acid extraction system based on magnetic separation. The design and implementation of the system are analyzed and investigated in-depth, focusing on the core methods, hardware control, and software control of the automated nucleic acid extraction system. Additionally, a study and evaluation were carried out concerning the nucleic acid extraction and detection aspects encompassed by the system. The results demonstrate that the temperature deviation in the lysis and elution fluids is approximately ±1 °C, the positioning accuracy of the system’s movement is ±0.005 mm, the average magnetic bead recovery rate is 94.98%, and the average nucleic acid recovery rate is 91.83%. The developed automated system and manual methods are employed for sample extraction, enabling the isolation of highly pure nucleic acids from bacteria, blood, and animal tissues for RT-PCR detection. The instrument employs lysis temperatures ranging from 70–80 °C, elution temperature of 80 °C, and drying time of 5–10 min, with a total extraction time of less than 35 min for different sample types. Overall, the system yields high nucleic acid concentration and purity, exhibits stable instrument operation, good repeatability, high efficiency, and low cost. It meets the requirements of genetic-level research and is worthy of clinical promotion and usage. Full article

(This article belongs to the Special Issue Functional Nanomaterials for Biosensing)

► Show Figures

Figure 1

17 pages, 2803 KiB

Open AccessArticle

Mechanistic Elucidation of Nanomaterial-Enhanced First-Generation Biosensors Using Probe Voltammetry of an Enzymatic Reaction

by Ann H. Wemple, Jamie S. Kaplan and Michael C. Leopold

Biosensors 2023, 13(8), 798; https://doi.org/10.3390/bios13080798 - 09 Aug 2023

Viewed by 899

Abstract

The incorporation of nanomaterials (NMs) into biosensing schemes is a well-established strategy for gaining signal enhancement. With electrochemical biosensors, the enhanced performance achieved from using NMs is often attributed to the specific physical properties of the chosen nanocomponents, such as their high electronic [...] Read more.

The incorporation of nanomaterials (NMs) into biosensing schemes is a well-established strategy for gaining signal enhancement. With electrochemical biosensors, the enhanced performance achieved from using NMs is often attributed to the specific physical properties of the chosen nanocomponents, such as their high electronic conductivity, size-dependent functionality, and/or higher effective surface-to-volume ratios. First generation amperometric biosensing schemes, typically utilizing NMs in conjunction with immobilized enzyme and semi-permeable membranes, can possess complex sensing mechanisms that are difficult to study and challenging to understand beyond the observable signal enhancement. This study shows the use of an enzymatic reaction between xanthine (XAN) and xanthine oxidase (XOx), involving multiple electroactive species, as an electrochemical redox probe tool for ascertaining mechanistic information at and within the modified electrodes used as biosensors. Redox probing using components of this enzymatic reaction are demonstrated on two oft-employed biosensing approaches and commonly used NMs for modified electrodes: gold nanoparticle doped films and carbon nanotube interfaces. In both situations, the XAN metabolism voltammetry allows for a greater understanding of the functionality of the semipermeable membranes, the role of the NMs, and how the interplay between the two components creates signal enhancement. Full article

(This article belongs to the Special Issue Functional Nanomaterials for Biosensing)

► Show Figures

Figure 1

10 pages, 2268 KiB

Open AccessCommunication

Biological Recognition-Based Electrochemical Aptasensor for Point-of-Care Detection of cTnI

by Jianfeng Ma, Lin Feng, Jie Li, Dan Zhu, Lianhui Wang and Shao Su

Biosensors 2023, 13(7), 746; https://doi.org/10.3390/bios13070746 - 19 Jul 2023

Cited by 1 | Viewed by 1429

Abstract

As a “gold standard biomarker”, cardiac troponin I (cTnI) is widely used to diagnose acute myocardial infarction (AMI). For an early clinical diagnosis of AMI, it is necessary to develop a facile, fast and on-site device for cTnI detection. According to this demand, [...] Read more.

As a “gold standard biomarker”, cardiac troponin I (cTnI) is widely used to diagnose acute myocardial infarction (AMI). For an early clinical diagnosis of AMI, it is necessary to develop a facile, fast and on-site device for cTnI detection. According to this demand, a point-of-care electrochemical aptasensor was developed for cTnI detection by coupling the advantages of screen-printed carbon electrode (SPCE) with those of an aptamer. Thiol and methylene blue (MB) co-labelled aptamer (MB-Apt-SH) was assembled on the surface of hierarchical flower-like gold nanostructure (HFGNs)-decorated SPCE (SPCE-HFGNs) to recognize and analyze cTnI. In the presence of cTnI, the specific biological recognition reaction between cTnI and aptamer caused the decrease in electrochemical signal. Under the optimal condition, this designed aptasensor showed wide linear range (10 pg/mL–100 ng/mL) and low detection limit for (8.46 pg/mL) for cTnI detection with high selectivity and stability. More importantly, we used a mobile phone coupled with a simple APP to efficiently detect cTnI in 10 μL 100% human serum samples, proving that this aptasensor has a promising potential in point-of-care testing. Full article

(This article belongs to the Special Issue Functional Nanomaterials for Biosensing)

► Show Figures

Figure 1

11 pages, 7750 KiB

Open AccessCommunication

Single-Atom Fe Nanozyme with Enhanced Oxidase-like Activity for the Colorimetric Detection of Ascorbic Acid and Glutathione

by Yue Gu, Zhongxu Cao, Mengde Zhao, Yanan Xu and Na Lu

Biosensors 2023, 13(4), 487; https://doi.org/10.3390/bios13040487 - 18 Apr 2023

Cited by 8 | Viewed by 2205

Abstract

Single-atom nanozymes (SAzymes) have drawn ever-increasing attention due to their maximum atom utilization efficiency and enhanced enzyme-like activity. Herein, a facile pyrolysis strategy is reported for the synthesis of the iron–nitrogen–carbon (Fe-N-C) SAzyme using ferrocene trapped within porous zeolitic imidazolate framework-8 (ZIF-8@Fc) as [...] Read more.

Single-atom nanozymes (SAzymes) have drawn ever-increasing attention due to their maximum atom utilization efficiency and enhanced enzyme-like activity. Herein, a facile pyrolysis strategy is reported for the synthesis of the iron–nitrogen–carbon (Fe-N-C) SAzyme using ferrocene trapped within porous zeolitic imidazolate framework-8 (ZIF-8@Fc) as a precursor. The as-prepared Fe-N-C SAzyme exhibited exceptional oxidase-mimicking activity, catalytically oxidizing 3,3′,5,5′-tetramethylbenzidine (TMB) with high affinity (K_m) and fast reaction rate (V_max). Taking advantage of this property, we designed two colorimetric sensing assays based on different interaction modes between small molecules and Fe active sites. Firstly, utilizing the reduction activity of ascorbic acid (AA) toward oxidized TMB (TMBox), a colorimetric bioassay for AA detection was established, which exhibited a good linear range of detection from 0.1 to 2 μM and a detection limit as low as 0.1 μM. Additionally, based on the inhibition of nanozyme activity by the thiols of glutathione (GSH), a colorimetric biosensor for GSH detection was constructed, showing a linear response over a concentration range of 1–10 μM, with a detection limit of 1.3 μM. This work provides a promising strategy for rationally designing oxidase-like SAzymes and broadening their application in biosensing. Full article

(This article belongs to the Special Issue Functional Nanomaterials for Biosensing)

► Show Figures

Figure 1

12 pages, 2707 KiB

Open AccessArticle

Label-Free Detection of T4 Polynucleotide Kinase Activity and Inhibition via Malachite Green Aptamer Generated from Ligation-Triggered Transcription

by Jingyi Si, Wei Zhou, Ying Fang, Da Zhou, Yifan Gao, Qunyan Yao, Xizhong Shen and Changfeng Zhu

Biosensors 2023, 13(4), 449; https://doi.org/10.3390/bios13040449 - 31 Mar 2023

Cited by 1 | Viewed by 1640

Abstract

Polynucleotide kinase (PNK) is a key enzyme that is necessary for ligation-based DNA repair. The activity assay and inhibitor screening for PNK may contribute to the prediction and improvement of tumor treatment sensitivity, respectively. Herein, we developed a simple, low-background, and label-free method [...] Read more.

Polynucleotide kinase (PNK) is a key enzyme that is necessary for ligation-based DNA repair. The activity assay and inhibitor screening for PNK may contribute to the prediction and improvement of tumor treatment sensitivity, respectively. Herein, we developed a simple, low-background, and label-free method for both T4 PNK activity detection and inhibitor screening by combining a designed ligation-triggered T7 transcriptional amplification system and a crafty light-up malachite green aptamer. Moreover, this method successfully detected PNK activity in the complex biological matrix with satisfactory outcomes, indicating its great potential in clinical practice. Full article

(This article belongs to the Special Issue Functional Nanomaterials for Biosensing)

► Show Figures

Figure 1

12 pages, 2351 KiB

Open AccessArticle

A Schematic Colorimetric Assay for Sialic Acid Assay Based on PEG-Mediated Interparticle Crosslinking Aggregation of Gold Nanoparticles

by Shixing Tang, Lin Li, Rui Wang, Sagar Regmi, Xinyu Zhang, Guoqiang Yang and Jian Ju

Biosensors 2023, 13(2), 164; https://doi.org/10.3390/bios13020164 - 20 Jan 2023

Cited by 2 | Viewed by 1983

Abstract

Sialic acid (SA) is a well-known component of glycoproteins, which have applications in various functional processes on the cell’s surface. The colorimetric is a simpler and more convenient method for measuring SA due to its low-cost apparatus and visual signal changes. This work [...] Read more.

Sialic acid (SA) is a well-known component of glycoproteins, which have applications in various functional processes on the cell’s surface. The colorimetric is a simpler and more convenient method for measuring SA due to its low-cost apparatus and visual signal changes. This work focused on the unpredictable interparticle crosslinking aggregation of the functionalized gold nanoparticles (AuNPs) in complex media. We proposed a balance of the Derjaguin–Landau–Verwey–Overbeek (DLVO)-type aggregation and molecule-based interaction method to solve this problem. Here, we report a novel colorimetric assay for the determination of SA using 4-mercaptophenyl boronic acid (4-MPBA) as an analyte’s recognition molecule, and negative charge PEG₄₀₀ was used to repulsive the interparticle crosslinking. The proposed sensing platform shows a linear relationship between the ratio of the absorbance intensity (A₅₂₅/A₆₆₀) and concentration of SA from 0.05 to 8 mM (R² = 0.997) and a detection limit of 48 μM was observed. The novel gold-based colorimetric sensor is easy to fabricate, reproducible in its test performance and has been successfully applied for the detection of SA in biological and healthcare product samples. Full article

(This article belongs to the Special Issue Functional Nanomaterials for Biosensing)

► Show Figures

Figure 1

15 pages, 3176 KiB

Open AccessArticle

Peptide Nanosheet-Inspired Biomimetic Synthesis of CuS Nanoparticles on Ti₃C₂ Nanosheets for Electrochemical Biosensing of Hydrogen Peroxide

by Danzhu Zhu, Hao Kong, Guozheng Yang, Peng He, Xin Luan, Lei Guo and Gang Wei

Biosensors 2023, 13(1), 14; https://doi.org/10.3390/bios13010014 - 22 Dec 2022

Cited by 2 | Viewed by 1711

Abstract

Hydrogen peroxide (H₂O₂) is one of the intermediates or final products of biological metabolism and participates in many important biological processes of life activities. The detection of H₂O₂ is of great significance in clinical disease monitoring, [...] Read more.

Hydrogen peroxide (H₂O₂) is one of the intermediates or final products of biological metabolism and participates in many important biological processes of life activities. The detection of H₂O₂ is of great significance in clinical disease monitoring, environmental protection, and bioanalysis. In this study, Ti₃C₂-based nanohybrids are prepared by the biological modification and self-assembled peptide nanosheets (PNSs)-based biomimetic synthesis of copper sulfide nanoparticles (CuS NPs), which show potential application in the fabrication of low-cost and high-performance electrochemical H₂O₂ biosensors. The synthesized CuS-PNSs/Ti₃C₂ nanohybrids exhibit excellent electrochemical performance towards H₂O₂, in which CuS NPs can catalyze the decomposition of H₂O₂ and realize the transformation from a chemical signal to an electrical signal to achieve the purpose of H₂O₂ detection. The prepared CuS-PNSs/Ti₃C₂-based electrochemical biosensor platform exhibits a wide detection range (5 μM–15 mM) and a low detection limit (0.226 μM). In addition, it reveals good selectivity and stability and can realize the monitoring of H₂O₂ in a complex environment. The successful biomimetic synthesis of CuS-PNSs/Ti₃C₂ hybrid nanomaterials provides a green and friendly strategy for the design and synthesis of functional nanomaterials and also provides a new inspiration for the construction of highly effective electrochemical biosensors for practical detection of H₂O₂ in various environments. Full article

(This article belongs to the Special Issue Functional Nanomaterials for Biosensing)

► Show Figures

Figure 1

9 pages, 2068 KiB

Open AccessArticle

Procedural Data Processing for Single-Molecule Identification by Nanopore Sensors

by Yupeng Wang, Jianxuan Yuan, Haofeng Deng, Ziang Zhang, Qianli D. Y. Ma, Lingzhi Wu and Lixing Weng

Biosensors 2022, 12(12), 1152; https://doi.org/10.3390/bios12121152 - 09 Dec 2022

Viewed by 1553

Abstract

Nanopores are promising single-molecule sensing devices that have been successfully used for DNA sequencing, protein identification, as well as virus/particles detection. It is important to understand and characterize the current pulses collected by nanopore sensors, which imply the associated information of the analytes, [...] Read more.

Nanopores are promising single-molecule sensing devices that have been successfully used for DNA sequencing, protein identification, as well as virus/particles detection. It is important to understand and characterize the current pulses collected by nanopore sensors, which imply the associated information of the analytes, including the size, structure, and surface charge. Therefore, a signal processing program, based on the MATLAB platform, was designed to characterize the ionic current signals of nanopore measurements. In a movable data window, the selected current segment was analyzed by the adaptive thresholds and corrected by multi-functions to reduce the noise obstruction of pulse signals. Accordingly, a set of single molecular events was identified, and the abundant information of current signals with the dwell time, amplitude, and current pulse area was exported for quantitative analysis. The program contributes to the efficient and fast processing of nanopore signals with a high signal-to-noise ratio, which promotes the development of the nanopore sensing devices in various fields of diagnosis systems and precision medicine. Full article

(This article belongs to the Special Issue Functional Nanomaterials for Biosensing)

► Show Figures

Figure 1

16 pages, 4905 KiB

Open AccessArticle

Bare Eye Detection of Bacterial Enzymes of Pseudomonas aeruginosa with Polymer Modified Nanoporous Silicon Rugate Filters

by Qasim Alhusaini, Walter Sebastian Scheld, Zhiyuan Jia, Dipankar Das, Faria Afzal, Mareike Müller and Holger Schönherr

Biosensors 2022, 12(12), 1064; https://doi.org/10.3390/bios12121064 - 22 Nov 2022

Viewed by 1574

Abstract

The fabrication, characterization and application of a nanoporous Silicon Rugate Filter (pSiRF) loaded with an enzymatically degradable polymer is reported as a bare eye detection optical sensor for enzymes of pathogenic bacteria, which is devoid of any dyes. The nanopores of pSiRF were [...] Read more.

The fabrication, characterization and application of a nanoporous Silicon Rugate Filter (pSiRF) loaded with an enzymatically degradable polymer is reported as a bare eye detection optical sensor for enzymes of pathogenic bacteria, which is devoid of any dyes. The nanopores of pSiRF were filled with poly(lactic acid) (PLA), which, upon enzymatic degradation, resulted in a change in the effective refractive index of the pSiRF film, leading to a readily discernible color change of the sensor. The shifts in the characteristic fringe patterns before and after the enzymatic reaction were analyzed quantitatively by Reflectometric Interference Spectroscopy (RIfS) to estimate the apparent kinetics and its dependence on enzyme concentration. A clear color change from green to blue was observed by the bare eye after PLA degradation by proteinase K. Moreover, the color change was further confirmed in measurements in bacterial suspensions of the pathogen Pseudomonas aeruginosa (PAO1) as well as in situ in the corresponding bacterial supernatants. This study highlights the potential of the approach in point of care bacteria detection. Full article

(This article belongs to the Special Issue Functional Nanomaterials for Biosensing)

► Show Figures

Graphical abstract

10 pages, 2197 KiB

Open AccessCommunication

Application of Heat-Enhancement for Improving the Sensitivity of Quartz Crystal Microbalance

by Chenglong Song, Zhihao Ma, Chenglong Li, Hongxing Zhang, Zhiqiang Zhu and Jie Wang

Biosensors 2022, 12(8), 643; https://doi.org/10.3390/bios12080643 - 15 Aug 2022

Cited by 7 | Viewed by 1430

Abstract

The use of quartz crystal microbalance in trace mass detection is restricted by unsatisfactory sensitivity, especially in damping media, due to the worsening of the quality factor of the damping resonator. The enhancement of the sensor performance could be realized by increasing the [...] Read more.

The use of quartz crystal microbalance in trace mass detection is restricted by unsatisfactory sensitivity, especially in damping media, due to the worsening of the quality factor of the damping resonator. The enhancement of the sensor performance could be realized by increasing the innate resonant frequency of quartz oscillators. Herein, increased working temperature of QCM systems was proved to bring an enhancement of the original resonant frequency. In addition, the measurement of ion osmotic pressure, single layer formation and single nucleotide polymorphism (SNP) at different temperatures demonstrated that an increased working temperature could enhance the sensitivity and accuracy, suggesting a potential application in a series of trace detections. Full article

(This article belongs to the Special Issue Functional Nanomaterials for Biosensing)

► Show Figures

Figure 1

Review

Jump to: Research

30 pages, 4707 KiB

Open AccessReview

Stimulus-Responsive DNA Hydrogel Biosensors for Food Safety Detection

by Huiyuan Wang, Xinyu Wang, Keqiang Lai and Juan Yan

Biosensors 2023, 13(3), 320; https://doi.org/10.3390/bios13030320 - 24 Feb 2023

Cited by 8 | Viewed by 2477

Abstract

Food safety has always been a major global challenge to human health and the effective detection of harmful substances in food can reduce the risk to human health. However, the food industry has been plagued by a lack of effective and sensitive safety [...] Read more.

Food safety has always been a major global challenge to human health and the effective detection of harmful substances in food can reduce the risk to human health. However, the food industry has been plagued by a lack of effective and sensitive safety monitoring methods due to the tension between the cost and effectiveness of monitoring. DNA-based hydrogels combine the advantages of biocompatibility, programmability, the molecular recognition of DNA molecules, and the hydrophilicity of hydrogels, making them a hotspot in the research field of new nanomaterials. The stimulus response property greatly broadens the function and application range of DNA hydrogel. In recent years, DNA hydrogels based on stimulus-responsive mechanisms have been widely applied in the field of biosensing for the detection of a variety of target substances, including various food contaminants. In this review, we describe the recent advances in the preparation of stimuli-responsive DNA hydrogels, highlighting the progress of its application in food safety detection. Finally, we also discuss the challenges and future application of stimulus-responsive DNA hydrogels. Full article

(This article belongs to the Special Issue Functional Nanomaterials for Biosensing)

► Show Figures

Figure 1

Journal Menu

Journal Browser

Functional Nanomaterials for Biosensing

Share This Special Issue

Special Issue Editor

Special Issue Information

Keywords

Published Papers (11 papers)

Research

Review

Further Information

Guidelines

MDPI Initiatives

Follow MDPI