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Recent Advance in Biosensors and Its Applications in Point-of-Care Molecular...
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Recent Advance in Biosensors and Its Applications in Point-of-Care Molecular Diagnostics (POC-MDx)

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

Deadline for manuscript submissions: closed (30 June 2024) | Viewed by 13792

Special Issue Editors

Dr. Kevin Eboigbodin

E-Mail Website
Guest Editor
Senior Director, Molecular Diagnostic Reagents, Medix Biochemica Group, Klovinpellontie 3, FI-02180 Espoo, Finland
Interests: molecular; diagnostics; point-of-care; microarrays; Infection; biomarkers; cancer diagnostics; virology; molecular biology; biochemistry; analytical chemistry; bioanalysis; medical diagnostics; precision medicine
Special Issues, Collections and Topics in MDPI journals
Dr. Luigi La Spada

E-Mail Website
Guest Editor
Lecturer in Electrical and Electronic Engineering, School of Engineering and the Built Environment, Edinburgh Napier University, 10 Colinton Rd, Edinburgh EH10 5DT, UK
Interests: metamaterials; metasurfaces; near-zero index materials; sensors; medical diagnostics; electromagnetics; sound waves; heat waves; fluid dynamics; classical mechanics
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Molecular diagnostics (MDx) is a collection of techniques that are used for the detection or monitoring of specific biomarkers in the genome and proteome levels. These techniques include, but are not limited to, polymerase chain reaction/real-time polymerase chain reaction (PCR/qPCR), isothermal nucleic acid amplification (iNAAT), whole genome sequencing (WGS), next-generation sequencing (NGS), microarrays, and clustered regularly interspaced short palindromic repeats (CRISPR). These techniques are typically performed within centralized and specialized testing laboratories. However, the increasing number of complex diseases, antibiotic-resistant pathogens as well as the recent outbreak of COVID-19 infections worldwide have further emphasized the need for more accurate, timely, and robust molecular diagnostic tests that can be used for self-testing or at the Point-of-Care (POC). Point-of-Care Molecular Diagnostics (POC-MDx) uses portable instruments or instrument free platforms and does not require specialized training to perform the test, thus enabling timely and accurate diagnosis outside the centralized laboratories. The use of POC-MDx tests will continue to grow rapidly due to the need for more personalized treatment and decentralized testing, representing a paradigm-shift in global healthcare.

Biosensors are an integral part of molecular diagnostics including POC-MDx test devices. The biosensors within the MDx device enable the detection or monitoring of specific biomarkers by converting their biological response into visual, electrical, or measurable signals. Monitoring specific biomarkers such as nucleic acids (DNA/RNA), enzymes, antibodies, proteins, and metabolites can provide information about metabolic disorders, disease state or changes to environmental conditions. Consequently, advances in biosensor technologies will also lead to advances in molecular diagnostics and POC applications. Advances in biosensor research and development in areas such as (but not limited to) visual detection, electrochemical detection, electromagnetic, optical, immunosensors, lab-chip, nanotechnology to bioelectronics, fluorescence detection, and nucleic acid biosensors will enable more robust and cost-effective MDx/POC-MDx tests. In this Special Issue, we welcome all research that helps to advance biosensors technologies and their application in molecular diagnostics.

Dr. Kevin Eboigbodin
Dr. Luigi La Spada
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

  • molecular diagnostics
  • biosensor
  • DNA, RNA
  • biomarker
  • point-of-care
  • home testing
  • self-testing
  • PCR, NGS
  • CRISPR
  • isothermal nucleic acid amplification

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Published Papers (5 papers)

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Research

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15 pages, 3010 KiB  
Article
Enhanced Point-of-Care SARS-CoV-2 Detection: Integrating RT-LAMP with Microscanning
by Minkyeong Choi, Eunji Lee, Seoyeon Park, Chae-Seung Lim and Woong-Sik Jang
Biosensors 2024, 14(7), 348; https://doi.org/10.3390/bios14070348 - 17 Jul 2024
Viewed by 1014
Abstract
The COVID-19 pandemic has highlighted the urgent need for rapid and accurate diagnostic methods for various infectious diseases, including SARS-CoV-2. Traditional RT-PCR methods, while highly sensitive and specific, require complex equipment and skilled personnel. In response, we developed an integrated RT-LAMP-MS assay, which [...] Read more.
The COVID-19 pandemic has highlighted the urgent need for rapid and accurate diagnostic methods for various infectious diseases, including SARS-CoV-2. Traditional RT-PCR methods, while highly sensitive and specific, require complex equipment and skilled personnel. In response, we developed an integrated RT-LAMP-MS assay, which combines rapid reverse transcription loop-mediated isothermal amplification (RT-LAMP) with microscanning (MS) technology for detecting SARS-CoV-2. The assay uses magnesium pyrophosphate formed during LAMP amplification as a visual marker, allowing direct observation via microscopy without the need for additional chemical indicators or probes. For the SARS-CoV-2/IC RT-LAMP-MS assay, the sample-LAMP reagent mixture was added to a microchip with SARS-CoV-2 primers and internal controls, then incubated at 62 °C for 30 min in a heat block, followed by amplification analysis using a microscanner. In clinical tests, the RT-LAMP-MS assay showed 99% sensitivity and 100% specificity, which is identical to the RT-LAMP results and comparable to the commercial AllplexTM SARS-CoV-2 assay results. Additionally, the limit of detection (LOD) was determined to be 10−1 PFU mL−1 (dynamic range: 103~10−1 PFU mL−1). The assay delivers results in 30 min, uses low-cost equipment, and demonstrates 100% reproducibility in repeated tests, making it suitable for point-of-care use in resource-limited settings. Full article
(This article belongs to the Special Issue Recent Advance in Biosensors and Its Applications in Point-of-Care Molecular Diagnostics (POC-MDx))
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15 pages, 3694 KiB  
Article
Detection of Reverse Transcriptase LAMP-Amplified Nucleic Acid from Oropharyngeal Viral Swab Samples Using Biotinylated DNA Probes through a Lateral Flow Assay
by Saloni Agarwal, Mojdeh Hamidizadeh and Frank F. Bier
Biosensors 2023, 13(11), 988; https://doi.org/10.3390/bios13110988 - 17 Nov 2023
Cited by 1 | Viewed by 2144
Abstract
This study focuses on three key aspects: (a) crude throat swab samples in a viral transport medium (VTM) as templates for RT-LAMP reactions; (b) a biotinylated DNA probe with enhanced specificity for LFA readouts; and (c) a digital semi-quantification of LFA readouts. Throat [...] Read more.
This study focuses on three key aspects: (a) crude throat swab samples in a viral transport medium (VTM) as templates for RT-LAMP reactions; (b) a biotinylated DNA probe with enhanced specificity for LFA readouts; and (c) a digital semi-quantification of LFA readouts. Throat swab samples from SARS-CoV-2 positive and negative patients were used in their crude (no cleaning or pre-treatment) forms for the RT-LAMP reaction. The samples were heat-inactivated but not treated for any kind of nucleic acid extraction or purification. The RT-LAMP (20 min processing time) product was read out by an LFA approach using two labels: FITC and biotin. FITC was enzymatically incorporated into the RT-LAMP amplicon with the LF-LAMP primer, and biotin was introduced using biotinylated DNA probes, specifically for the amplicon region after RT-LAMP amplification. This assay setup with biotinylated DNA probe-based LFA readouts of the RT-LAMP amplicon was 98.11% sensitive and 96.15% specific. The LFA result was further analysed by a smartphone-based IVD device, wherein the T-line intensity was recorded. The LFA T-line intensity was then correlated with the qRT-PCR Ct value of the positive swab samples. A digital semi-quantification of RT-LAMP-LFA was reported with a correlation coefficient of R2 = 0.702. The overall RT-LAMP-LFA assay time was recorded to be 35 min with a LoD of three RNA copies/µL (Ct-33). With these three advancements, the nucleic acid testing-point of care technique (NAT-POCT) is exemplified as a versatile biosensor platform with great potential and applicability for the detection of pathogens without the need for sample storage, transportation, or pre-processing. Full article
(This article belongs to the Special Issue Recent Advance in Biosensors and Its Applications in Point-of-Care Molecular Diagnostics (POC-MDx))
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13 pages, 6058 KiB  
Communication
Surface-Enhanced Raman Scattering Active Core-Shell Ag NPs@Carbon Dots with Enzyme-Mimicking Activities for Label-Free Measurement Cholesterol
by Jian Ju, Lin Li, Bei Li, Sagar Regmi, Tingting Wang, Jiao Xu, Chaojie Li and Shixing Tang
Biosensors 2023, 13(10), 927; https://doi.org/10.3390/bios13100927 - 16 Oct 2023
Viewed by 2025
Abstract
Serological-sensitive testing of cholesterol holds significant value in the fields of healthcare and clinical diagnosis. This study reports on the preparation of peroxidase-mimicking nanozymes through the wrapping of N, S-doped carbon dots (DCDs) on the surface of silver nanoparticles (Ag NPs@DCD). The shell–core [...] Read more.
Serological-sensitive testing of cholesterol holds significant value in the fields of healthcare and clinical diagnosis. This study reports on the preparation of peroxidase-mimicking nanozymes through the wrapping of N, S-doped carbon dots (DCDs) on the surface of silver nanoparticles (Ag NPs@DCD). The shell–core structure of Ag NPs@DCD displays peroxidase-mimicking capability, with the potential to catalyze inactive Raman probe molecules into the Raman reporters. Furthermore, a “shell-isolated nanoparticles-enhanced Raman spectroscopy” structure exhibited an enhanced Raman signal of reporter molecules. Ag NPs@DCD were utilized to create a label-free SERS sensing system for high-performance detection of cholesterol in serum samples. These results demonstrate the potential of the novel nanozyme-based SERS approach for clinical diagnosis. Full article
(This article belongs to the Special Issue Recent Advance in Biosensors and Its Applications in Point-of-Care Molecular Diagnostics (POC-MDx))
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Review

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21 pages, 1474 KiB  
Review
Detection of Parasites in the Field: The Ever-Innovating CRISPR/Cas12a
by Xin Li, Zhisheng Dang, Wenqiang Tang, Haoji Zhang, Jianwei Shao, Rui Jiang, Xu Zhang and Fuqiang Huang
Biosensors 2024, 14(3), 145; https://doi.org/10.3390/bios14030145 - 14 Mar 2024
Cited by 1 | Viewed by 3070
Abstract
The rapid and accurate identification of parasites is crucial for prompt therapeutic intervention in parasitosis and effective epidemiological surveillance. For accurate and effective clinical diagnosis, it is imperative to develop a nucleic-acid-based diagnostic tool that combines the sensitivity and specificity of nucleic acid [...] Read more.
The rapid and accurate identification of parasites is crucial for prompt therapeutic intervention in parasitosis and effective epidemiological surveillance. For accurate and effective clinical diagnosis, it is imperative to develop a nucleic-acid-based diagnostic tool that combines the sensitivity and specificity of nucleic acid amplification tests (NAATs) with the speed, cost-effectiveness, and convenience of isothermal amplification methods. A new nucleic acid detection method, utilizing the clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) nuclease, holds promise in point-of-care testing (POCT). CRISPR/Cas12a is presently employed for the detection of Plasmodium falciparum, Toxoplasma gondii, Schistosoma haematobium, and other parasites in blood, urine, or feces. Compared to traditional assays, the CRISPR assay has demonstrated notable advantages, including comparable sensitivity and specificity, simple observation of reaction results, easy and stable transportation conditions, and low equipment dependence. However, a common issue arises as both amplification and cis-cleavage compete in one-pot assays, leading to an extended reaction time. The use of suboptimal crRNA, light-activated crRNA, and spatial separation can potentially weaken or entirely eliminate the competition between amplification and cis-cleavage. This could lead to enhanced sensitivity and reduced reaction times in one-pot assays. Nevertheless, higher costs and complex pre-test genome extraction have hindered the popularization of CRISPR/Cas12a in POCT. Full article
(This article belongs to the Special Issue Recent Advance in Biosensors and Its Applications in Point-of-Care Molecular Diagnostics (POC-MDx))
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21 pages, 8218 KiB  
Review
Advances in Point-of-Care Testing of microRNAs Based on Portable Instruments and Visual Detection
by Zhong-Yu Wang, Ming-Hui Sun, Qun Zhang, Pei-Feng Li, Kun Wang and Xin-Min Li
Biosensors 2023, 13(7), 747; https://doi.org/10.3390/bios13070747 - 20 Jul 2023
Cited by 9 | Viewed by 2895
Abstract
MicroRNAs (miRNAs) are a class of small noncoding RNAs that are approximately 22 nt in length and regulate gene expression post-transcriptionally. miRNAs play a vital role in both physiological and pathological processes and are regarded as promising biomarkers for cancer, cardiovascular diseases, neurodegenerative [...] Read more.
MicroRNAs (miRNAs) are a class of small noncoding RNAs that are approximately 22 nt in length and regulate gene expression post-transcriptionally. miRNAs play a vital role in both physiological and pathological processes and are regarded as promising biomarkers for cancer, cardiovascular diseases, neurodegenerative diseases, and so on. Accurate detection of miRNA expression level in clinical samples is important for miRNA-guided diagnostics. However, the common miRNA detection approaches like RNA sequencing, qRT-PCR, and miRNA microarray are performed in a professional laboratory with complex intermediate steps and are time-consuming and costly, challenging the miRNA-guided diagnostics. Hence, sensitive, highly specific, rapid, and easy-to-use detection of miRNAs is crucial for clinical diagnosis based on miRNAs. With the advantages of being specific, sensitive, efficient, cost-saving, and easy to operate, point-of-care testing (POCT) has been widely used in the detection of miRNAs. For the first time, we mainly focus on summarizing the research progress in POCT of miRNAs based on portable instruments and visual readout methods. As widely available pocket-size portable instruments and visual detection play important roles in POCT, we provide an all-sided discussion of the principles of these methods and their main limitations and challenges, in order to provide a guide for the development of more accurate, specific, and sensitive POCT methods for miRNA detection. Full article
(This article belongs to the Special Issue Recent Advance in Biosensors and Its Applications in Point-of-Care Molecular Diagnostics (POC-MDx))
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