Detection Of Disease Biomarkers Research Articles

A novel signal amplified sandwich aptamer-based visual lateral flow assay based on self-assembly bifunctional nucleic acid for rapid and sensitive detection of CKMB

At present, rapid and sensitive detection of disease biomarkers is still the focus of early clinical diagnosis. As a rapid response to point-of-care testing (POCT), lateral flow assay (LFA) has been given serious attention. Unfortunately, due to the sensitivity limitations, LFA, especially AuNPs-based LFA, was difficult to play an important role in the detection of trace biomarkers. The combination of different signal amplification methods increases the complexity of operation and use, or prolongs the detection time, which sacrifices the advantages of fast and convenient of LFA. Hence, a novel sandwich aptamer-based visual LFA based on bifunctional nucleic acid (bFNA-LFA) was proposed for rapid and sensitive detection of creatine kinase MB (CKMB) in this study. Based on hybridization chain reaction (HCR) and nucleic acid hybridization, a bifunctional nucleic acid with target recognition and signal amplification was prepared, which effectively improved the sensitivity of CKMB detection without increasing the reaction time and depending on the detecting equipment. Under optimal reaction conditions, the limit of detection of 7.92 ng/mL was observed, which was 20 times lower than that of sandwich aptamer-based AuNPs LFA in the previous study. The proposed bFNA-LFA assay expanded a new idea in improving the sensitivity of LFA, and provided a reference for the subsequent application of nucleic acid nanomaterials in the detection of protein biomarkers.

A novel method for multiple cardiovascular disease biomarker detection using a SERF-based microfluidic platform

As cardiovascular diseases (CVDs) are the leading cause of death worldwide, it is of great importance to realize a rapid and sensitive diagnosis. Multiple CVD biomarker detection, which holds higher accuracy and speed of early CVD diagnosis, is becoming a promising alternative to single biomarker analysis. Cardiac troponin I (cTnI), C-reactive protein (CRP), brain-type natriuretic peptide (BNP), and heart fatty acid binding protein (H-FABP) are four significant diagnostic biomarkers for CVDs. Among the several methods for biomarker detection, such as optical, magnetic, mechanical, and electrical methods, the magnetic method stands out because of its stability, low toxicity, noninvasive detection, facile manipulation with an external magnetic field, and immunity to the interference of background light. At present, the lack of multiplexing capability, the imprecision of magnetic signal localization, and the agglomeration and non-specific adsorption of magnetic nanoparticles all limit the clinical application of magnetic detection. This paper presented a novel method for multiple CVD biomarker detection using a spin exchange relaxation-free (SERF)-based microfluidic platform. One or more SERF atomic magnetometers were used to scan the remanent field of magnetic labels to detect four CVD biomarkers in 70min, with a detection limit of 1.88pg/mL (cTnI), 1.58pg/mL (CRP), 1.45pg/mL (BNP), and 1.71pg/mL (H-FABP). The results showed two-dimensional magnetic fields with a resolution of submillimeter level and a sensitivity of pT level. The measurement technique provides a new magnetic methodology for applications in clinical diagnosis.

Enzymes as indispensable markers in disease diagnosis.

Enzymes have been usedfor disease diagnosis for many decades; however, advancements in technology like ELISA and flow cytometry-based detection havesignificantlyincreased their use and have increased the sensitivity of detection. Technological advancements in recombinant enzyme production haveincreased enzymatic stability,and the use ofcolorimetric-based and florescence-based assays has led to their increased use as biomarkers for disease detection. Enzymes like acid phosphatase, cathepsin, lactate dehydrogenase, thymidine kinaseand creatine kinase are indispensable markers for diagnosing cancer, cardiovasculardiseasesand others. This minireview summarizes various enzymes used in disease diagnosis, their metabolic role, market valueand potential as disease markers across various metabolic and other disorders.

FeNi-MIL-88B-based electrochemiluminescence immunosensor for ultra-sensitive detection of CD44 protein via dual-quenching strategy

BackgroundCluster of Differentiation 44 (CD44) is considered an important biomarker for various cancers, and achieving highly sensitive detection of CD44 is crucial, which plays a significant role in tumor invasion and metastasis, providing essential information for clinical tumor diagnosis. Commonly used methods for analysis include fluorescence spectroscopy (FL), photoelectrochemical analysis (PEC), electrochemical analysis (EC), and commercial ELISA kits. Although these methods offer high sensitivity, they can be relatively complex to perform experimentally. Electrochemiluminescence (ECL) has gained widespread research attention due to its high sensitivity, ease of operation, effective spatiotemporal control, and close to zero background signal. ResultsIn this work, a sandwich-type ECL immunosensor for detecting CD44 was constructed using luminol as a luminophore. In this sensing platform, bimetallic MOFs (Pd@FeNi-MIL-88B) loaded with palladium nanoparticles (Pd NPs) were used as a novel enzyme mimic, exhibiting excellent catalytic performance towards the electroreduction of H2O2. The hybrids provided a strong support platform for luminol and antibodies, significantly enhancing the initial ECL signal of luminol. Subsequently, core-shell Au@MnO2 nanocomposites were synthesised by gold nanoparticles (Au NPs) encapsulated in manganese dioxide (MnO2) thin layers, as labels. In the luminol/H2O2 system, Au@MnO2 exhibited strong light absorption in the broad UV–vis spectrum, similar to the black body effect, and the scavenging effect of Mn2+ on O2•−, which achieved the dual-quenching of ECL signal. Under the optimal experimental conditions, the immunosensor demonstrated a detection range of 0.1 pg mL−1 − 100 ng mL−1, with a detection limit of 0.069 pg mL−1. SignificanceBased on Pd@FeNi-MIL-88B nanoenzymes and Au@MnO2 nanocomposites, a dual-quenching sandwich-type ECL immunosensor for the detection of CD44 was constructed. The proposed immunosensor exhibited excellent reproducibility, stability, selectivity, and sensitivity, and provided a valuable analytical strategy and technical platform for the accurate detection of disease biomarkers, and opened up potential application prospects for early clinical treatment.

Metal-enhanced fluorescence biosensor integrated in capillary flow-driven microfluidic cartridge for highly sensitive immunoassays

Point-of-care testing (POCT) for low-concentration protein biomarkers remains challenging due to limitations in biosensor sensitivity and platform integration. This study addresses this gap by presenting a novel approach that integrates a metal-enhanced fluorescence (MEF) biosensor within a capillary flow-driven microfluidic cartridge (CFMC) for the ultrasensitive detection of the Parkinson's disease biomarker, aminoacyl-tRNA synthetase complex interacting multi-functional protein 2 (AIMP-2). Crucial point to this approach is the orientation-controlled immobilization of capture antibody on a nanodimple-structured MEF substrate within the CFMC. This strategy significantly enhances fluorescence signals without quenching, enabling accurate quantification of low-concentration AIMP-2 using a simple digital fluorescence microscope with a light-emitting diode excitation source and a digital camera. The resulting platform exhibits exceptional sensitivity, achieving a limit of detection in the pg/mL range for AIMP-2 in human serum. Additionally, the CFMC design incorporates a capillary-driven passive sample transport mechanism, eliminating the need for external pumps and further simplifying the detection process. Overall, this work demonstrates the successful integration of MEF biosensing with capillary microfluidics for point-of-care applications.

Coupled Poly(ethylenimine) Coreactant to Enhance Electrochemiluminescence of Polymer Dots for Array Imaging of Protein Biomarkers.

Traditional electrochemiluminescent (ECL) bioanalysis suffers from the demand for excessive external coreactants and the damage of reaction intermediates. In this work, a poly(ethylenimine) (PEI)-coupled ECL emitter was proposed by covalently coupling tertiary amine-rich PEI to polymer dots (Pdots). The coupled PEI might act as a highly efficient coreactant to enhance the ECL emission of Pdots through intramolecular electron transfer, reducing the electron transfer distance between emitter and coreactant intermediates and avoiding the disadvantages of traditional ECL systems. Through modification of the PEI-Pdots with tDNA, a sequence partially complementary to cDNA that was complementary to the aptamer of target protein biomarker (aDNA), tDNA-PEI-Pdots were obtained. The biosensors were produced using Au/indium tin oxide (ITO) with an aDNA/cDNA hybrid, and an ECL imaging biosensor array was constructed for ultrasensitive detection of protein biomarkers. Using vascular endothelial growth factor 165 (VEGF165) as a protein model, the proposed ECL imaging method containing two simple incubations with target samples and then tDNA-PEI-Pdots showed a detectable range of 1 pg mL-1 to 100 ng mL-1 and a detection limit of 0.71 pg mL-1, as well as excellent performance such as low toxicity, high sensitivity, excellent selectivity, good accuracy, and acceptable fabrication reproducibility. The PEI-coupled Pdots provide a new avenue for the design of ECL emitters and the application of ECL imaging in disease biomarker detection.

Understanding Circular RNAs in Health, Welfare, and Productive Traits of Cattle, Goats, and Sheep.

Circular RNAs (circRNAs) are unique noncoding RNA molecules, notable for their covalent closed-loop structures, which play a crucial role in regulating gene expression across a variety of biological processes. This review comprehensively synthesizes the existing knowledge of circRNAs in three key livestock species: Bos taurus (cattle), Ovis aries (sheep), and Capra hircus (goats). It focuses on their functional importance and emerging potential as biomarkers for disease detection, stress response, and overall physiological health. Specifically, it delves into the expression and functionality of circRNAs in these species, paying special attention to traits critical to livestock productivity such as milk production, meat quality, muscle development, wool production, immune responses, etc. We also address the current challenges faced in circRNA research, including the need for standardized methodologies and broader studies. By providing insights into the molecular mechanisms regulated by circRNAs, this review underscores their scientific and economic relevance in the livestock industry. The potential of circRNAs to improve animal health management and the quality of animal-derived products aligns with growing consumer concerns for animal welfare and sustainability. Thus, this paper aims to guide future research directions while supporting the development of innovative strategies in livestock management and breeding.

Recent Advances in Biosensors for Diagnosis of Autoimmune Diseases.

Over the last decade, autoimmune diseases (ADs) have undergone a significant increase because of genetic and/or environmental factors; therefore, their simple and fast diagnosis is of high importance. The conventional diagnostic techniques for ADs require tedious sample preparation, sophisticated instruments, a dedicated laboratory, and qualified personnel. For these reasons, biosensors could represent a useful alternative to these methods. Biosensors are considered to be promising tools that can be used in clinical analysis for an early diagnosis due to their high sensitivity, simplicity, low cost, possible miniaturization (POCT), and potential ability for real-time analysis. In this review, recently developed biosensors for the detection of autoimmune disease biomarkers are discussed. In the first part, we focus on the main AD biomarkers and the current methods of their detection. Then, we discuss the principles and different types of biosensors. Finally, we overview the characteristics of biosensors based on different bioreceptors reported in the literature.

Seamless Integration of Rapid Separation and Ultrasensitive Detection for Complex Biological Samples Using Multistage Annular Functionalized Carbon Nanotube Arrays.

Efficient separation, enrichment, and detection of bacteria in diverse media are pivotal for identifying bacterial diseases and their transmission pathways. However, conventional bacterial detection methods that split the separation and detection steps are plagued by prolonged processing times. Herein, a multistage annular functionalized carbon nanotube array device designed for the seamless integration of complex biological sample separation and multimarker detection is introduced. This device resorts to the supersmooth fluidity of the liquid sample in the carbon nanotubes interstice through rotation assistance, achieving the ability to quickly separate impurities and capture biomarkers (1mL sample cost time of 2.5 s). Fluid dynamics simulations show that the reduction of near-surface hydrodynamic resistance drives the capture of bacteria and related biomarkers on the functionalized surface of carbon nanotube in sufficient time. When further assembled as an even detection device, it exhibited fast detection (<30min), robust linear correlation (101 -107 colony-forming units [CFU] mL-1 , R2 = 0.997), ultrasensitivity (limit of detection = 1.7 CFU mL-1 ), and multitarget detection (Staphylococcus aureus, extracellular vesicles, and enterotoxin proteins). Collectively, the material and system offer an expanded platform for real-time diagnostics, enabling integrated rapid separation and detection of various disease biomarkers.

Assessment of Cellulose Nanofiber-Based Elastase Biosensors to Inflammatory Disease as a Function of Spacer Length and Fluorescence Response.

Inflammatory disease biomarker detection has become a high priority in point-of-care diagnostic research in relation to chronic wounds, with a variety of sensor-based designs becoming available. Herein, two primary aspects of biosensor design are examined: (1) assessment of a cellulose nanofiber (CNF) matrix derived from cotton ginning byproducts as a sensor transducer surface; and (2) assessment of the relation of spacer length and morphology between the CNF cellulose backbone and peptide fluorophore as a function of sensor activity for porcine pancreatic and human neutrophil elastases. X-ray crystallography, specific surface area, and pore size analyses confirmed the suitability of CNF as a matrix for wound care diagnostics. Based upon the normalized degree of substitution, a pegylated-linker connecting CNF transducer substrate to peptide fluorophore showed the greatest fluorescence response, compared to short- and long-chain alkylated linkers.

A Point‐of‐Care Sensing Platform for Multiplexed Detection of Chronic Kidney Disease Biomarkers Using Molecularly Imprinted Polymers

AbstractChronic kidney disease (CKD) is one of the most serious non‐communicable diseases affecting the population. In the early‐stages patients have no obvious symptoms, until it becomes life‐threatening leading end‐stage kidney failure. Therefore, it is important to early diagnose CKD to allow therapeutic interventions and progression monitoring. Here, a point‐of‐care (POC) sensing platform is reported for the simultaneous detection of three CKD biomarkers, namely creatinine, urea, and human serum albumin (HSA), using reduced graphene oxide/polydopamine‐molecularly imprinted polymer (rGO/PDA‐MIP) fabricated with novel surface‐molecularly imprinting technology. A multi‐channel electrochemical POC readout system with differential pulse voltammetry (DPV) function is developed, allowing the simultaneous detection of the three biomarkers, in combination with the surface‐MIP electrodes. This sensing platform achieves the record low limit‐of‐detection (LoD) at a femtomolar level for all three analytes, with wide detection ranges covering their physiological concentrations. Clinical validation is performed by measuring these analytes in serum and urine from healthy controls and patients with CKD. The average recovery rate is 81.8–119.1% compared to the results obtained from the hospital, while this platform is more cost‐effective, user‐friendly, and requires less sample‐to‐result time, showing the potential to be deployed in resource‐limited settings for the early diagnosis and tracking progression of CKD.

Anti‐Fouling Polymer or Peptide‐Modified Electrochemical Biosensors for Improved Biosensing in Complex Media

AbstractElectrochemical biosensing represents a highly effective technology for detecting disease biomarkers given its high sensitivity, low and clinically relevant limit of detection, and cost effectiveness. However, in complex media such as urine, blood, sweat or saliva, biosensing performance can be significantly impacted by electrode biofouling by proteins, cells, lipids, and other matrix components. Such biofouling leads to reduced signal from the target analyte coupled with an elevated background signal, resulting in poor signal‐to‐noise ratios (SNRs), reduced sensitivity, and lower specificity. This comprehensive review describes the design of anti‐fouling polymers and peptides as a potential solution to prevent or suppress electrochemical biosensor fouling. Various anti‐fouling polymers and peptides developed for improved biosensing in complex media are summarized in the context of their mechanism(s) of anti‐fouling, methods of deposition, and practical applications. Recent advances and persistent challenges in the field are also reviewed to provide perspectives on new directions toward enhancing anti‐fouling in electrochemical biosensors.

Ultrafast, high-accuracy disease biomarker detection through fluorogenic single-molecule recognition

Ultrafast, high-accuracy disease biomarker detection through fluorogenic single-molecule recognition

Electrochemical peptide-based biosensor for the detection of the inflammatory disease biomarker, interleukin-1beta

This paper reports the development of a highly sensitive and selective electrochemical peptide-based biosensor for the detection of the inflammatory disease biomarker, interleukin-1beta (IL-1β). To this end, flower-like Au–Ag@MoS2-rGO nanocomposites were used as the signal amplification platform to achieve a label-free biosensor with a high sensitivity and selectivity. First, a high-affinity peptide for IL-1β was identified through biopanning with M13 random peptide libraries, and was newly designed by incorporating cysteine at the C-terminus. An IL-1β specific binding peptide was used as the bio-receptor, and the interaction between the IL-1β binding peptide and IL-1β was confirmed via enzyme-linked immunosorbent assay and various physicochemical and electrochemical analyses. Under optimal conditions, the biosensor achieved an ultrasensitive and specific IL-1β detection in a wide linear concentration range of 0–250 ng/mL with a picomolar-level detection limit (∼2.4 pM), low binding constant (∼0.62 pM), and a low coefficient of variation (<1.65 %). The biosensor was successfully utilized for IL-1β determination in the serum of Crohn's disease patients with a good correlation coefficient. In addition, the detection performance was comparable to that of commercially available IL-1β ELISA kit. This indicates that the electrochemical peptide-based biosensor may offer a potentially valuable platform for the clinical diagnosis of various inflammatory disease biomarkers.

Electrochemically Synthesized MIP Sensors: Applications in Healthcare Diagnostics.

Early-stage detection and diagnosis of diseases is essential to the prompt commencement of treatment regimens, curbing the spread of the disease, and improving human health. Thus, the accurate detection of disease biomarkers through the development of robust, sensitive, and selective diagnostic tools has remained cutting-edge scientific research for decades. Due to their merits of being selective, stable, simple, and having a low preparation cost, molecularly imprinted polymers (MIPs) are increasingly becoming artificial substitutes for natural receptors in the design of state-of-the-art sensing devices. While there are different MIP preparation approaches, electrochemical synthesis presents a unique and outstanding method for chemical sensing applications, allowing the direct formation of the polymer on the transducer as well as simplicity in tuning the film properties, thus accelerating the trend in the design of commercial MIP-based sensors. This review evaluates recent achievements in the applications of electrosynthesized MIP sensors for clinical analysis of disease biomarkers, identifying major trends and highlighting interesting perspectives on the realization of commercial MIP-endowed testing devices for rapid determination of prevailing diseases.

Shining a light on liver health: advancements in fluorescence-enhanced enzyme biosensors for early disease detection.

Early detection of liver diseases holds paramount importance in optimizing treatment outcomes and prognosis, thereby significantly enhancing the likelihood of recovery while mitigating the risk of progression to liver cancer. Liver diseases encompass a spectrum of conditions, each potentially manifesting distinct enzymatic profiles. Monitoring these enzymes in situ facilitates timely intervention and therapeutic management. In recent years, the field of biosensor technology has witnessed remarkable advancement, owing to strides in biomedicine and computational sciences. Biosensors have garnered widespread utility across medical and biological domains, spanning the detection of disease biomarkers, drug release tracking, ion imaging, and fluorescence imaging within living organisms. These applications have markedly enhanced imaging resolution and have the potential to refine disease diagnosis accuracy for clinicians. A pivotal aspect in the successful application of this technology lies in the construction of fluorescence probes adept at swiftly and selectively identifying target enzymes by amalgamating liver disease enzymes with fluorescence probe technology. However, research in this niche area remains relatively scarce. Building upon this foundational understanding, the present review delineates the utilization of biosensors in the early diagnosis of liver disease. Serving as a theoretical framework, this review envisages the development of high-performance biosensors tailored for the early detection of liver cancer. Furthermore, it offers insights into the potential of biosensor technology to progress and broaden its practical applications, thus contributing to the advancement of diagnostic methodologies in liver disease management.

Multiplexed electrical detection of whole viruses from plasma in a microfluidic platform.

The advancement of point-of-care diagnostics is crucial to improving patient outcomes, especially in areas with low access to hospitals or specialized laboratories. In particular, rapid, sensitive, and multiplexed detection of disease biomarkers has great potential to achieve accurate diagnosis and inform high quality care for patients. Our Coulter counting and immunocapture based detection system has previously shown its broad applicability in the detection of cells, proteins, and nucleic acids. This paper expands the capability of the platform by demonstrating multiplexed detection of whole-virus particles using electrically distinguishable hydrogel beads by demonstrating the capability of our platform to achieve simultaneous detection at clinically relevant concentrations of hepatitis A virus (>2 × 103 IU mL-1) and human parvovirus B19 virus like particles (>106 IU mL-1) from plasma samples. The expanded versatility of the differential electrical counting platform allows for more robust and diverse testing capabilities.

Construction of a streptavidin-based dual-localized DNAzyme walker for disease biomarker detection.

A dual-localized DNAzyme walker (dlDW) was constructed by utilizing multiple split DNAzymes with probes, and their substrates are separately localized on streptavidin and AuNPs, serving as walking pedals and tracks, respectively. Based on dlDW, biosensing platform was successfully constructed and showed great potential application in clinical disease diagnosis.

One-Component Dual-Readout Aggregation-Induced Emission Nanobeads for Qualitative and Quantitative Detection of C-Reactive Protein at the Point of Care.

Fluorescent lateral flow immunoassay (LFA) systems are versatile tools for sensitive and quantitative detection of disease markers at the point of care. However, traditional fluorescent nanoparticle-based lateral flow immunoassays are not visible under room light, necessitate an additional fluorescent reader, and lack flexibility for different application scenarios. Herein, we report a dual-readout LFA system for the rapid and sensitive detection of C-reactive protein (CRP) in clinical samples. The system relied on the aggregation-induced emission nanobeads (AIENBs) encapsulated with red AIE luminogen, which possesses both highly fluorescent and colorimetric properties. The AIENB-based LFA in the naked-eye mode was able to qualitatively detect CRP levels as low as 8.0 mg/L, while in the fluorescent mode, it was able to quantitatively measure high-sensitivity CRP (hs-CRP) with a limit of detection of 0.16 mg/L. The AIENB-based LFA system also showed a good correlation with the clinically used immunoturbidimetric method for CRP and hs-CRP detection in human plasma. This dual-modal AIENB-based LFA system offers the convenience of colorimetric testing and highly sensitive and quantitative detection of disease biomarkers and medical diagnostics in various scenarios.

Fluorescent imaging and smartphone detection of ClO- using sensors based on trifluoromethylquinoline-substituted pyranonitrile

In the field of novel fluorescent sensors, achieving rapid and sensitive tracking of target substances has become an urgent imperative. Applying this technology to the detection of biomarkers for biological diseases holds profound practical significance. Hypochlorous acid (HClO) is a significant reactive oxygen species (ROS) within biological systems. Aberrant metabolism of HClO can result in disruptions of physiological functions and oxidative stress, consequently influencing the development of various diseases. In this context, our study has successfully developed a novel fluorescent probe named W-4a, which employs trifluoromethyl quinoline as a substitute for traditional pyranonitrile compounds, enabling the rapid (<40 s) and highly sensitive (UV limit of detection: 1.62 μM; fluorescence limit of detection: 0.14 μM) detection of ClO-, during the detection process, ClO- effectively activated the fluorescent probe and showed a distinct red fluorescent signal at 687 nm. Furthermore, the W-4a probe exhibits excellent biocompatibility and excels in both exogenous and endogenous ClO- detection imaging within cells, Co-localization experiments indicate that probe W-4a effectively targets the Golgi apparatus. Additionally, in conjunction with the W-4a probe, we have devised a portable smart detection method for hypochlorous acid in environmental settings, which holds the potential to serve as a flexible and potent tool for monitoring environmental toxicity.