Research

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16 pages, 3830 KiB

Open AccessArticle

Enhancing Glucose Biosensing with Graphene Oxide and Ferrocene-Modified Linear Poly(ethylenimine)

by Jirawan Monkrathok, Pattanaphong Janphuang, Somphong Suphachiaraphan, Sastiya Kampaengsri, Anyanee Kamkaew, Kantapat Chansaenpak, Sireerat Lisnund, Vincent Blay and Piyanut Pinyou

Biosensors 2024, 14(4), 161; https://doi.org/10.3390/bios14040161 - 28 Mar 2024

Viewed by 582

Abstract

We designed and optimized a glucose biosensor system based on a screen-printed electrode modified with the NAD-GDH enzyme. To enhance the electroactive surface area and improve the electron transfer efficiency, we introduced graphene oxide (GO) and ferrocene-modified linear poly(ethylenimine) (LPEI-Fc) onto the biosensor [...] Read more.

We designed and optimized a glucose biosensor system based on a screen-printed electrode modified with the NAD-GDH enzyme. To enhance the electroactive surface area and improve the electron transfer efficiency, we introduced graphene oxide (GO) and ferrocene-modified linear poly(ethylenimine) (LPEI-Fc) onto the biosensor surface. This strategic modification exploits the electrostatic interaction between graphene oxide, which possesses a negative charge, and LPEI-Fc, which is positively charged. This interaction results in increased catalytic current during glucose oxidation and helps improve the overall glucose detection sensitivity by amperometry. We integrated the developed glucose sensor into a flow injection (FI) system. This integration facilitates a swift and reproducible detection of glucose, and it also mitigates the risk of contamination during the analyses. The incorporation of an FI system improves the efficiency of the biosensor, ensuring precise and reliable results in a short time. The proposed sensor was operated at a constant applied potential of 0.35 V. After optimizing the system, a linear calibration curve was obtained for the concentration range of 1.0–40 mM (R² = 0.986). The FI system was successfully applied to determine the glucose content of a commercial sports drink. Full article

(This article belongs to the Special Issue Biosensing for Point-of-Care Diagnostics)

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15 pages, 4638 KiB

Open AccessArticle

A Chamber-Based Digital PCR Based on a Microfluidic Chip for the Absolute Quantification and Analysis of KRAS Mutation

by Jie Ren, Gangwei Xu, Hongna Liu, Nongyue He, Zhehao Zhao, Meiling Wang, Peipei Gu, Zhu Chen, Yan Deng, Dongping Wu and Song Li

Biosensors 2023, 13(8), 778; https://doi.org/10.3390/bios13080778 - 01 Aug 2023

Viewed by 1329

Abstract

The Kirsten rat sarcoma virus gene (KRAS) is the most common tumor in human cancer, and KRAS plays an important role in the growth of tumor cells. Normal KRAS inhibits tumor cell growth. When mutated, it will continuously stimulate cell growth, resulting in [...] Read more.

The Kirsten rat sarcoma virus gene (KRAS) is the most common tumor in human cancer, and KRAS plays an important role in the growth of tumor cells. Normal KRAS inhibits tumor cell growth. When mutated, it will continuously stimulate cell growth, resulting in tumor development. There are currently few drugs that target the KRAS gene. Here, we developed a microfluidic chip. The chip design uses parallel fluid channels combined with cylindrical chamber arrays to generate 20,000 cylindrical microchambers. The microfluidic chip designed by us can be used for the microsegmentation of KRAS gene samples. The thermal cycling required for the PCR stage is performed on a flat-panel instrument and detected using a four-color fluorescence system. “Glass-PDMS-glass” sandwich structure effectively reduces reagent volatilization; in addition, a valve is installed at the sample inlet and outlet on the upper layer of the chip to facilitate automatic control. The liquid separation performance of the chip was verified by an automated platform. Finally, using the constructed KRAS gene mutation detection system, it is verified that the chip has good application potential for digital polymerase chain reaction (dPCR). The experimental results show that the chip has a stable performance and can achieve a dynamic detection range of four orders of magnitude and a gene mutation detection of 0.2%. In addition, the four-color fluorescence detection system developed based on the chip can distinguish three different KRAS gene mutation types simultaneously on a single chip. Full article

(This article belongs to the Special Issue Biosensing for Point-of-Care Diagnostics)

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15 pages, 4555 KiB

Open AccessArticle

A New Optical Interferometric Biosensing System Enhanced with Nanoparticles for Alzheimer’s Disease in Serum

by Ana María M. Murillo, María Fe Laguna, Luis G. Valle, Luca Tramarin, Yolanda Ramirez, Álvaro Lavín, Beatriz Santamaría and Miguel Holgado

Biosensors 2023, 13(7), 707; https://doi.org/10.3390/bios13070707 - 05 Jul 2023

Cited by 3 | Viewed by 1472

Abstract

In this scientific work, we demonstrate, for the first time, a new biosensing system and procedure to measure specifically the total Tau (T-Tau) protein in serum, one of the most relevant biomarkers of Alzheimer’s disease (AD). AD is a progressive brain disorder that [...] Read more.

In this scientific work, we demonstrate, for the first time, a new biosensing system and procedure to measure specifically the total Tau (T-Tau) protein in serum, one of the most relevant biomarkers of Alzheimer’s disease (AD). AD is a progressive brain disorder that produces neuronal and cognitive dysfunction and affects a high percentage of people worldwide. For this reason, diagnosing AD at the earliest possible stage involves improving diagnostic systems. We report on the use of interferometric bio-transducers integrated with 65 microwells forming diagnostic KITs read-out by using the Interferometric Optical Detection Method (IODM). Moreover, biofunctionalized silicon dioxide (SiO₂) nanoparticles (NPs) acting as interferometric enhancers of the bio-transducers signal allow for the improvement of both the optical read-out signal and its ability to work with less-invasive biological samples such as serum instead of cerebrospinal fluid (CSF). As a result, in this paper, we describe for the first time a relevant diagnostic alternative to detect Tau protein at demanding concentrations of 10 pg/mL or even better, opening the opportunity to be used for detecting other relevant AD-related biomarkers in serum, such as β-amyloid and phosphorylated Tau (P-Tau), neurofilaments, among others that can be considered relevant for AD. Full article

(This article belongs to the Special Issue Biosensing for Point-of-Care Diagnostics)

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15 pages, 2560 KiB

Open AccessArticle

Surface Micro-Patterned Biofunctionalized Hydrogel for Direct Nucleic Acid Hybridization Detection

by Paola Zezza, María Isabel Lucío, Estrella Fernández, Ángel Maquieira and María-José Bañuls

Biosensors 2023, 13(3), 312; https://doi.org/10.3390/bios13030312 - 23 Feb 2023

Cited by 3 | Viewed by 1667

Abstract

The present research is focused on the development of a biofunctionalized hydrogel with a surface diffractive micropattern as a label-free biosensing platform. The biosensors described in this paper were fabricated with a holographic recording of polyethylene terephthalate (PET) surface micro-structures, which were then [...] Read more.

The present research is focused on the development of a biofunctionalized hydrogel with a surface diffractive micropattern as a label-free biosensing platform. The biosensors described in this paper were fabricated with a holographic recording of polyethylene terephthalate (PET) surface micro-structures, which were then transferred into a hydrogel material. Acrylamide-based hydrogels were obtained with free radical polymerization, and propargyl acrylate was added as a comonomer, which allowed for covalent immobilization of thiolated oligonucleotide probes into the hydrogel network, via thiol-yne photoclick chemistry. The comonomer was shown to significantly contribute to the immobilization of the probes based on fluorescence imaging. Two different immobilization approaches were demonstrated: during or after hydrogel synthesis. The second approach showed better loading capacity of the bioreceptor groups. Diffraction efficiency measurements of hydrogel gratings at 532 nm showed a selective response reaching a limit of detection in the complementary DNA strand of 2.47 µM. The label-free biosensor as designed could significantly contribute to direct and accurate analysis in medical diagnosis as it is cheap, easy to fabricate, and works without the need for further reagents. Full article

(This article belongs to the Special Issue Biosensing for Point-of-Care Diagnostics)

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12 pages, 1357 KiB

Open AccessArticle

A Microfluidic Platform with an Embedded Miniaturized Electrochemical Sensor for On-Chip Plasma Extraction Followed by In Situ High-Sensitivity C-Reactive Protein (hs-CRP) Detection

by Zhi-Xuan Lai, Chia-Chien Wu and Nien-Tsu Huang

Biosensors 2022, 12(12), 1163; https://doi.org/10.3390/bios12121163 - 13 Dec 2022

Cited by 3 | Viewed by 2091

Abstract

Blood testing is a clinical diagnostic tool to evaluate physiological conditions, the immune system response, or the presence of infection from whole blood samples. Although conventional blood testing can provide rich biological information, it usually requires complicated and tedious whole blood processing steps [...] Read more.

Blood testing is a clinical diagnostic tool to evaluate physiological conditions, the immune system response, or the presence of infection from whole blood samples. Although conventional blood testing can provide rich biological information, it usually requires complicated and tedious whole blood processing steps operated by benchtop instruments and well-experienced technicians, limiting its usage in point-of-care (POC) settings. To address the above problems, we propose a microfluidic platform for on-chip plasma extraction directly from whole blood and in situ biomarker detection. Herein, we chose C-reactive protein (CRP) as the target biomarker, which can be used to predict fatal cardiovascular disease (CVD) events such as heart attacks and strokes. To achieve a rapid, undiluted, and high-purity on-chip plasma extraction, we combined two whole blood processing methods: (1) anti-D immunoglobulin-assisted sedimentation, and (2) membrane filtration. To perform in situ CRP detection, we fabricated a three-dimensional (3D) microchannel with an embedded electrochemical (EC) sensor, which has a modular design to attach the blood collector and buffer reservoir with standard Luer connectors. As a proof of concept, we first confirmed that the dual plasma extraction design achieved the same purity level as the standard centrifugation method with smaller sample (100 µL of plasma extracted from 400 µL of whole blood) and time (7 min) requirements. Next, we validated the functionalization protocol of the EC sensor, followed by evaluating the detection of CRP spiked in plasma and whole blood. Our microfluidic platform performed on-chip plasma extraction directly from whole blood and in situ CRP detection at a 0.1–10 μg/mL concentration range, covering the CVD risk evaluation level of the high-sensitivity CRP (hs-CRP) test. Based on the above features, we believe that this platform constitutes a flexible way to integrate the processing of complex samples with accurate biomarker detection in a sample-to-answer POC platform, which can be applied in CVD risk monitoring under critical clinical situations. Full article

(This article belongs to the Special Issue Biosensing for Point-of-Care Diagnostics)

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11 pages, 1426 KiB

Open AccessArticle

A Lateral Flow Device for Point-of-Care Detection of Doxorubicin

by Tania Pomili, Francesca Gatto and Pier Paolo Pompa

Biosensors 2022, 12(10), 896; https://doi.org/10.3390/bios12100896 - 19 Oct 2022

Cited by 4 | Viewed by 1722

Abstract

A simple, rapid, and sensitive point-of-care (POC) device for the on-site detection of doxorubicin was developed. The proposed method relies on the naked-eye detection of the intrinsic fluorescence of the drug in a lateral flow device (LFD) configuration, exploiting the biological recognition of [...] Read more.

A simple, rapid, and sensitive point-of-care (POC) device for the on-site detection of doxorubicin was developed. The proposed method relies on the naked-eye detection of the intrinsic fluorescence of the drug in a lateral flow device (LFD) configuration, exploiting the biological recognition of DNA probes and avoiding the use of expensive antibodies and sophisticated instrumentations. The POC assay does not require any pre-treatment or purification step and provides an immediate visual readout, achieving a limit of detection as low as ca. 1 ng doxorubicin, outperforming several laboratory-based instrumental techniques. The POC method was proven useful for the detection of trace amounts of the drug both in the case of water solutions (to simulate surface contaminations) and in urine samples, opening promising perspectives for routine monitoring of doxorubicin, with potential benefit to healthcare workers and personalized chemotherapies. Full article

(This article belongs to the Special Issue Biosensing for Point-of-Care Diagnostics)

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Review

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14 pages, 9532 KiB

Open AccessReview

Last Trends in Point-of-Care (POC) Diagnostics for the Management of Hematological Indices in Home Care Patients

by Fabrizio Clemente, Amina Antonacci, Maria Teresa Giardi, Valeria Frisulli, Francesco Paolo Tambaro and Viviana Scognamiglio

Biosensors 2023, 13(3), 345; https://doi.org/10.3390/bios13030345 - 04 Mar 2023

Cited by 4 | Viewed by 2130

Abstract

Today, complete blood count (CBC) analyses are highly automated and allow for high throughput and accurate and reliable results. However, new analytical tools are in great demand to provide simple, rapid and cost-effective management of hematological indices in home care patients. Chronic disease [...] Read more.

Today, complete blood count (CBC) analyses are highly automated and allow for high throughput and accurate and reliable results. However, new analytical tools are in great demand to provide simple, rapid and cost-effective management of hematological indices in home care patients. Chronic disease monitoring at home has become a benefit for patients who are finding cost savings in programs designed to monitor/treat patients in offsite locations. This review reports the latest trends in point-of-care (POC) diagnostics useful for home testing of key hematological counts that may be affected during home therapy treatment. Full article

(This article belongs to the Special Issue Biosensing for Point-of-Care Diagnostics)

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32 pages, 6811 KiB

Open AccessReview

A review of Optical Point-of-Care devices to Estimate the Technology Transfer of These Cutting-Edge Technologies

by María Jesús Pioz, Rocío L. Espinosa, María Fe Laguna, Beatriz Santamaria, Ana María M. Murillo, Álvaro Lavín Hueros, Sergio Quintero, Luca Tramarin, Luis G Valle, Pedro Herreros, Alberto Bellido, Rafael Casquel and Miguel Holgado

Biosensors 2022, 12(12), 1091; https://doi.org/10.3390/bios12121091 - 29 Nov 2022

Cited by 5 | Viewed by 2759

Abstract

Despite the remarkable development related to Point-of-Care devices based on optical technology, their difficulties when used outside of research laboratories are notable. In this sense, it would be interesting to ask ourselves what the degree of transferability of the research work to the [...] Read more.

Despite the remarkable development related to Point-of-Care devices based on optical technology, their difficulties when used outside of research laboratories are notable. In this sense, it would be interesting to ask ourselves what the degree of transferability of the research work to the market is, for example, by analysing the relation between the scientific work developed and the registered one, through patent. In this work, we provide an overview of the state-of-the-art in the sector of optical Point-of-Care devices, not only in the research area but also regarding their transfer to market. To this end, we explored a methodology for searching articles and patents to obtain an indicator that relates to both. This figure of merit to estimate this transfer is based on classifying the relevant research articles in the area and the patents that have been generated from these ones. To delimit the scope of this study, we researched the results of a large enough number of publications in the period from 2015 to 2020, by using keywords “biosensor”, “optic”, and “device” to obtain the most representative articles from Web of Science and Scopus. Then, we classified them according to a particular classification of the optical PoC devices. Once we had this sampling frame, we defined a patent search strategy to cross-link the article with a registered patent (by surfing Google Patents) and classified them accordingly to the categories described. Finally, we proposed a relative figure called Index of Technology Transference (IoTT), which estimates to what extent our findings in science materialized in published articles are protected by patent. Full article

(This article belongs to the Special Issue Biosensing for Point-of-Care Diagnostics)

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27 pages, 12033 KiB

Open AccessReview

Recent Advances in Biosensor Technologies for Point-of-Care Urinalysis

by Chuljin Hwang, Won-June Lee, Su Dong Kim, Sungjun Park and Joo Hee Kim

Biosensors 2022, 12(11), 1020; https://doi.org/10.3390/bios12111020 - 15 Nov 2022

Cited by 11 | Viewed by 4825

Abstract

Human urine samples are non-invasive, readily available, and contain several components that can provide useful indicators of the health status of patients. Hence, urine is a desirable and important template to aid in the diagnosis of common clinical conditions. Conventional methods such as [...] Read more.

Human urine samples are non-invasive, readily available, and contain several components that can provide useful indicators of the health status of patients. Hence, urine is a desirable and important template to aid in the diagnosis of common clinical conditions. Conventional methods such as dipstick tests, urine culture, and urine microscopy are commonly used for urinalysis. Among them, the dipstick test is undoubtedly the most popular owing to its ease of use, low cost, and quick response. Despite these advantages, the dipstick test has limitations in terms of sensitivity, selectivity, reusability, and quantitative evaluation of diseases. Various biosensor technologies give it the potential for being developed into point-of-care (POC) applications by overcoming these limitations of the dipstick test. Here, we present a review of the biosensor technologies available to identify urine-based biomarkers that are typically detected by the dipstick test and discuss the present limitations and challenges that future development for their translation into POC applications for urinalysis. Full article

(This article belongs to the Special Issue Biosensing for Point-of-Care Diagnostics)

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