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Biosensors

Biosensors is an international, peer-reviewed, open access journal on the technology and science of biosensors, published monthly online by MDPI.

Indexed in PubMed | Quartile Ranking JCR - Q1 (Instruments and Instrumentation | Chemistry, Analytical)

All Articles (5,083)

Rapid Forensic DNA Profiling via Real-Time Recombinase Polymerase Amplification of InDel Markers

  • Liesl De Keyzer,
  • Sonja Škevin and
  • Filip Van Nieuwerburgh
  • + 2 authors

Forensic DNA profiling commonly relies on polymerase chain reaction (PCR) amplification followed by capillary electrophoresis (CE) or massively parallel sequencing (MPS), which requires expensive, laboratory-based equipment that depends on a stable power supply and is unsuitable for field applications. Here, we present a proof-of-concept assay that uses recombinase polymerase amplification (RPA) combined with exo probe detection for rapid, isothermal genotyping of insertion–deletion (InDel) markers. To the best of our knowledge, this study represents the first demonstration of forensic DNA typing using RPA coupled with exo probes. The reaction proceeds at 39 °C and combines amplification and detection in a single 20 min step. Thirteen DNA samples were genotyped in triplicate across eight InDel loci using allele-specific fluorescent probes. Genotypes were derived from differential endpoint fluorescence between matched and mismatched probes. Compared with benchmark genotyping, 97.07% of genotypes (n = 307) were correct at 1 ng DNA input. Accurate profiles were reliably obtained for DNA inputs as low as 250 pg, and partial profiles were still detectable at 31 pg. The results demonstrate that RPA-based InDel genotyping is fast, sensitive, and reproducible. With further optimization, such as refined probe design and selection of robust loci, the assay has clear potential to achieve complete accuracy and to be integrated into portable lab-on-a-chip platforms for rapid, field-deployable forensic identification.

6 February 2026

(A) Design of the InDel probes. InDel markers are highlighted in red. Each probe contains a tetrahydrofuran (THF) residue (red triangle), and is labelled with a FAM fluorophore (green) and a BHQ-1 quencher (black). Hybridization to the complementary allele in the presence of exonuclease III results in probe cleavage and fluorescence. Mismatched probes show reduced duplex stability and fluorescence intensity. (B) Illustration of a real-time RPA plot. Fluorescence is measured over time. Perfectly matched probes yield sigmoidal curves with higher endpoint values than mismatched probes.

Rare-earth-doped upconversion nanoparticles (UCNPs) exhibit upconversion luminescence upon excitation with infrared light and have been extensively utilized in the field of biosensing. In this study, a UCNPs-based biosensor with porous silicon (PSi) as the substrate was developed for the first time, enabling the detection of target DNA molecule concentration. First, a PSi substrate was prepared via electrochemical etching and subsequently functionalized to enable target DNA molecules to immobilize onto the inner walls of the PSi substrate’s pores. Then, UCNPs-labeled probe DNA molecules hybridized with the target DNA molecules, enabling indirect attachment of UCNPs to the inner walls of the PSi substrate. Subsequently, the sample surface is irradiated with a 980 nm laser. Upconversion fluorescence images of the sample, both before and after the biological reaction, are captured using an image acquisition device. Image processing software is employed to calculate the average change in grayscale values, enabling the determination of the molecular concentration of target DNA. The limit of detection (LOD) of this method for target DNA molecular concentration is 86 pM, demonstrating that it enables low-cost, highly sensitive, rapid, and convenient biological detection of target DNA molecules.

6 February 2026

SEM images of the PSi substrate: (a) surface SEM image; (b) cross-sectional SEM image.

Microfluidic chip technologies, also known as lab-on-a-chip systems, have profoundly transformed laboratory medicine by enabling the miniaturization, automation, and rapid processing of complex diagnostic assays using minimal sample volumes. Recent advances in chip design, fabrication methods—including 3D printing, modular and flexible substrates—and biosensor integration have significantly enhanced the performance, sensitivity, and clinical applicability of these devices. Integration of advanced biosensors allows for real-time detection of circulating tumor cells, nucleic acids, and exosomes, supporting innovative applications in cancer diagnostics, infectious disease detection, point-of-care testing (POCT), personalized medicine, and therapeutic monitoring. Notably, the convergence of microfluidics with artificial intelligence (AI) and machine learning has amplified device automation, reliability, and analytical power, resulting in “smart” diagnostic platforms capable of self-optimization, automated analysis, and clinical decision support. Emerging applications in fields such as neuroscience diagnostics and microbiome profiling further highlight the broad potential of microfluidic technology. Here, we present findings from a comprehensive review of recent innovations in microfluidic chip design and fabrication, advances in biosensor and AI integration, and their clinical applications in laboratory medicine. We also discuss current challenges in manufacturing, clinical validation, and system integration, as well as future directions for translating next-generation microfluidic technologies into routine clinical and public health practice.

5 February 2026

Overview of recent advances and clinical applications of microfluidic chip technologies in laboratory medicine. Microfluidic platforms are enabling breakthroughs across cancer diagnostics (e.g., CTC and ctDNA analysis, multi-omics liquid biopsy), point-of-care testing (including wearable POCT, cardiac and metabolic biomarker assays), therapeutic efficiency (such as tumor-on-chip drug screening and organs-on-chip immunotherapy), infectious disease detection (rapid pathogen identification, COVID-19 testing), neurological and therapeutic monitoring (brain-on-chip, real-time drug monitoring), and environmental health monitoring (genomics, proteomics, metabolomics, single-cell omics). Integrated detection modalities—including fluorescence, electrochemical sensors, flow cytometry (FCM), and mass spectrometry (MS)—alongside artificial intelligence (AI), further enhance microfluidic chip performance, automation, and data interpretation for personalized and precision medicine.

Sulfamethazine (SMZ) is widely used in livestock production, and its residues can enter water and soil environments, posing potential risks to human health and ecosystems. This study focuses on environmental samples and constructs an AuNP-based colorimetric aptasensor using the SMZ1S aptamer for the rapid visual detection of SMZ. Under optimized conditions, the aptasensor showed a wide linear range from 0.05 to 0.4 µg/mL and a limit of detection of 0.039 µg/mL. Molecular dynamics simulations have demonstrated that the aptamer’s binding to SMZ is stable, providing a theoretical basis for the high selectivity of the aptasensor. Spike-and-recovery experiments yielded recoveries of 87.3–105.5%, 88.6–102.8%, and 87.5–103.4% for SMZ in lake water, tap water, and soil samples, respectively, with relative standard deviations of 5.9–8.3%, 8.0–10.6%, and 4.8–9.6%, showing good agreement with high-performance liquid chromatography (HPLC) results (R2 ≥ 0.981). Overall, the proposed aptasensor provides a simple and effective approach for rapid detection of SMZ in environmental samples.

5 February 2026

Principle of aptasensor based on AuNPs for detection of SMZ.

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Sensors and Technology
Editors: Nélia Jordão Alberto, Maria de Fátima Domingues, Nunzio Cennamo, Adriana Borriello

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Biosensors - ISSN 2079-6374