Sensitivity Of Antigen Detection Research Articles

Ultrasensitive Specific Detection of Anti-influenza A H1N1 Hemagglutinin Monoclonal Antibody Using Silicon Nanowire Field Effect Biosensors.

Rapid and sensitive detection of virus-related antigens and antibodies is crucial for controlling sudden seasonal epidemics and monitoring neutralizing antibody levels after vaccination. However, conventional detection methods still face challenges related to compatibility with rapid, highly sensitive, and compact detection apparatus. In this work, we developed a Si nanowire (SiNW)-based field-effect biosensor by precisely controlling the process conditions to achieve the required electrical properties via complementary metal-oxide-semiconductor (CMOS)-compatible nanofabrication processes. The SiNW surface was chemically modified with 2-aminoethylphosphonic acid, followed by a dehydration condensation reaction with influenza A H1N1 hemagglutinin (HA1), to enable specific detection of anti-HA1 immunoglobulin G (IgG). We successfully detected the anti-influenza IgG with concentrations ranging from 1 aM to 100 nM, achieving a remarkable detection limit of 6.0 aM. To demonstrate specificity, a control experiment was conducted using normal mouse IgG with concentrations of 6 aM to 600 nM. The results showed a high specificity, with the signal being 6-fold greater for the target IgG compared to the control IgG. This work demonstrates the capability of SiNW biosensors to detect anti-influenza A H1N1 hemagglutinin monoclonal antibody with enhanced detection sensitivity and specificity. This work lays the groundwork for future applications in detecting antibodies after vaccination or immunotherapy, contributing to the effective management of infectious pandemics.

Crystal Structures of Antigen-Binding Fragment of Anti-Osteocalcin Antibody KTM219.

Osteocalcin is a useful biomarker for bone formation and bone-related diseases. KTM219 is an anti-osteocalcin C-terminal peptide antibody. The single-chain variable region (scFv) and antigen-binding fragment (Fab) of KTM219 are applicable to the Quenchbody (Q-body) immunoassay. Q-body is a new type of fluorescent immunosensor, which is scFv or Fab labeled with a fluorescent dye. When Q-body binds to its antigen, the fluorescence intensity increases. The highly sensitive detection of antigens by changes in fluorescence intensity is performed in a single step by mixing the sample and reagent. In this study, to reveal the recognition mechanism of the KTM219 antibody and to discuss the structural basis for Q-body, we solved the crystal structures of Fab of the anti-osteocalcin antibody KTM219 and its complex with the antigen osteocalcin C-terminal peptide (BGP-C7). Also, we solved the structure of a KTM219 Fab crystal grown in the presence of a fluorescent dye, carboxytetramethylrhodamine (TAMRA); however, tightly bound TAMRA was not found in the electron density map. We predicted the binding sites of TAMRA in the antigen-binding pocket by docking simulations. These results support the proposed Q-body mechanism. The crystal structures of KTM219 Fab would be useful for further development and improvement of Q-body fluorescent immunosensors.

Lattice atom-bridged chemical bond interface facilitates charge transfer for boosted photoelectric response

ABSTRACT The construction of chemical bonds at heterojunction interfaces currently presents a promising avenue for enhancing photogenerated carrier interfacial transfer. However, the deliberate modulation of these interfacial chemical bonds remains a significant challenge. In this study, we successfully established a p-n junction composed of atomic-level Pt-doped CeO2 and 2D metalloporphyrins metal-organic framework nanosheets (Pt-CeO2/CuTCPP(Fe)), which enables the realization of photoelectric enhancement by regulating the interfacial Fe–O bond and optimizing the built-in electric field. Atomic-level Pt doping in CeO2 leads to an increased density of oxygen vacancies and lattice mutation, which induces a transition in interfacial Fe–O bonds from adsorbed oxygen (Fe–OA) to lattice oxygen (Fe–OL). This transition changes the interfacial charge flow pathway from Fe–OA–Ce to Fe–OL, effectively reducing the carrier transport distance along the atomic-level charge transport highway. This results in a 2.5-fold enhancement in photoelectric performance compared with the CeO2/CuTCPP(Fe). Furthermore, leveraging the peroxidase-like activity of the p-n junction, we employed this functional heterojunction interface to develop a photoelectrochemical immunoassay for the sensitive detection of prostate-specific antigens.

Amino-functionalized vertically ordered mesoporous silica film on electrochemically polarized screen-printed carbon electrodes for the construction of gated electrochemical aptasensors and sensitive detection of carcinoembryonic antigens.

Disposable electrochemical biosensors with high sensitivity are very fit for point-of-care testing in clinical diagnosis. Herein, amino-functionalized, vertically ordered mesoporous silica films (NH2-VMSF) attached to an electrochemically polarized screen-printed carbon electrode (p-SPCE) are prepared using a simple electrochemical method and then utilized to construct a gated electrochemical aptasensor for rapid and sensitive determination of carcinoembryonic antigen (CEA). After being treated with the electrochemical polarization procedure, p-SPCE has plentiful oxygen-containing groups and improved catalytic ability, which help promote the stability of NH2-VMSF on SPCE without the use of an adhesive layer and simultaneously generate a highly electroactive sensing interface. Owing to the numerous uniform and ultrasmall nanopores of NH2-VMSF, CEA-specific aptamer anchored on the external surface of NH2-VMSF/p-SPCE serves as the gatekeeper, allowing the specific recognition and binding of CEA and eventually impeding the ingress of electrochemical probes [Fe(CN)6 3-/4-] through the silica nanochannels. The declined electrochemical responses of Fe(CN)6 3-/4- can be used to quantitatively detect CEA, yielding a wide detection range (100fg/mL to 100ng/mL) and a low limit of detection (24fg/mL). Moreover, the proposed NH2-VMSF/p-SPCE-based electrochemical aptasensor can be applied to detect the amount of CEA in spiked human serum samples, which extends the biological application of a disposable NH2-VMSF/p-SPCE sensor by modulating the biological recognition species.

An epitope imprinted electrochemical sensor for highly sensitive and selective determination of prostate-specific antigen.

Asimple method forhighly selective and sensitive prostate-specific antigen (PSA) detection using a molecularly imprinted electrochemical sensor ispresented. The sensor was developed through an epitope imprinted strategy combined with electrochemical measurement techniques. An epitope molecularly imprinted polymer (EMIP) film was constructed on a AuNPs-coated gold electrode surface through electropolymerization, utilizing the C-terminus epitope of PSA (KWIKDTIVANP) as the template molecular and o-phenylenediamine as the functional monomer. The characteristics of EMIP film were investigated by using a scanning electron microscope and electrochemical test methods, including electrochemical impedance spectroscopy and cyclic voltammetry. Key parameters such as electropolymerization cycles, elution and rebinding times, and the molar ratio of template molecular to functional monomer were systematically optimized. The sensor demonstrated a detection limit (LOD) of 0.31 fg/mL and exhibited an excellent linear response towards PSA concentration ranging from 1.0 fg/mL to 0.1 µg/mL. Furthermore, the EMIP sensor showed excellent selectivity against other biological macromolecules, such as bovine serum albumin, human serum albumin, alpha-fetoprotein, and carcinoembryonic antigen. With recoveriesbetween 95.89 and 106.04% for PSA detection in human serums theEMIP/AuNPs/AuE electrochemical sensor showed great potential in real sample analysis.

Integrating zirconia-gold hybrid into aptasensor capable of ultrasensitive and robust carcinoembryonic antigen determination

Aptasensors have emerged as a powerful strategy for clinical diagnostics due to their enhanced specificity and selectivity in detecting biomarkers. However, striking a delicate equilibrium between sensitivity, specificity, and stability remains a formidable challenge due to the biomarker's low abundance and clinical biospecimen's complexity. To address this, we developed a hybrid of zirconia-gold nanoparticles (ZrO2-AuNPs) to enhance the performance of aptasensors, resulting in improved sensitivity and stability. The introduction of ZrO2 resulted in a significant reduction in signal fluctuation, as demonstrated by a remarkable decrease in the relative standard deviation (RSD) from 22.60% to an impressive 1.95%, indicating improved stability; this enhancement was also observed in various ZrO2-AuNPs-aptasensors, yielding a minimal RSD of 2.61%. The sensitivity boosters (ZrO2 and AuNPs) amplified the electrochemical signal, leading to highly sensitive carcinoembryonic antigen (CEA) detection with a threshold as low as 42.504 fg/mL. For clinical application, the ZrO2-AuNPs-aptasensor demonstrated a strong correlation with clinical measurements, as evidenced by the regression equation y (A) = 1.005 × x - 0.025 (ng/mL) and a notable R2 value of 0.991, indicating a high predictive accuracy. Additionally, the aptasensor displayed remarkable diagnostic accuracy in distinguishing lung cancer from healthy samples, as evidenced by a perfect sensitivity of 1.0, a specificity of 0.913, and an impressive AUC value of 0.993 in the ROC analysis, a claim further confirmed by a significantly low p-value of 1.990 × 10-5 from a validation t-test. Moreover, we optimized various conditions, including temperature and pH, for the ZrO2-AuNPs-aptasensor, resulting in desirable reproducibility and selectivity. We anticipate that the ZrO2-AuNPs strategy will enhance the aptasensor's sensitivity and stability, potentially propelling advancements in clinical diagnostics.

Color-encoded multicompartmental hydrogel microspheres for multiplexed bioassays

We develop color-encoded multicompartmental hydrogel (MH) microspheres tailored for multiplexed bioassays using a drop-based microfluidic approach. Our method involves the creation of triple emulsion drops that feature thin sacrificial oil layers separating two prepolymer phases. This configuration leads to the formation of poly(ethylene glycol) (PEG) multi-compartmental core-shell microspheres through photopolymerization, followed by the removal of the thin oil layers. The core compartments stably incorporate pigments, ensuring their retention within the hydrogel network without leakage, which facilitates reliable color encoding across varying spatial positions. Additionally, we introduce small molecule fluorescent labeling into the chemically functionalized shell compartments, achieving consistent distribution of functional components without the core's contamination. Importantly, our integrated one-pot conjugation of these color-encoded microspheres with affinity peptides enables the highly sensitive and selective detection of influenza virus antigens using a fluorescence bioassay, resulting in an especially low detection limit of 0.18 nM and 0.66 nM for influenza virus H1N1 and H5N1 antigens, respectively. This approach not only highlights the potential of our microspheres in clinical diagnostics but also paves the way for their application in a wide range of multiplexed assays.

Sensitive Dual-Signal ELISA Based on Specific Phage-Displayed Double Peptide Probes with Internal Filtering Effect to Assay Monkeypox Virus Antigen.

The global spread of monkeypox has become a worldwide public healthcare issue. Therefore, there is an urgent need for accurate and sensitive detection methods to effectively control its spreading. Herein, we screened by phage display two peptides M4 (sequence: DPCGERICSIAL) and M6 (sequence: SCSSFLCSLKVG) with good affinity and specificity to monkeypox virus (MPXV) B21R protein. To simulate the state of the peptide in the phage and to avoid spatial obstacles of the peptide, GGGSK was added at the C terminus of M4 and named as M4a. Molecular docking shows that peptide M4a and peptide M6 are bound to different epitopes of B21R by hydrogen bonds and salt-bridge interactions, respectively. Then, peptide M4a was selected as the capture probe, phage M6 as the detection probe, and carbonized polymer dots (CPDs) as the fluorescent probe, and a colorimetric and fluorescent double-signal capture peptide/antigen/signal peptide-displayed phage sandwich ELISA triggered by horseradish peroxidase (HRP) through a simple internal filtration effect (IFE) was constructed. HRP catalyzes H2O2 to oxidize 3,3',5,5'-tetramethylbenzidine (TMB) to generate blue oxidized TMB, which can further quench the fluorescence of CPDs through IFE, enabling to detect MPXV B21R in colorimetric and fluorescent modes. The proposed simple immunoassay platform shows good sensitivity and reliability in MPXV B21R detection. The limit of detection for colorimetric and fluorescent modes was 27.8 and 9.14 pg/mL MPXV B21R, respectively. Thus, the established double-peptide sandwich-based dual-signal immunoassay provides guidance for the development of reliable and sensitive antigen detection capable of mutual confirmation, which also has great potential for exploring various analytical strategies for other respiratory virus surveillance.

Bio-inspired wettability patterning for flexible and wearable electrochemical immunosensors utilizing PDMS-TiO2 coating

The electrochemical sensor strip presented in this work, designed for low-volume point-of-care immunosensing applications and demonstrated using prostate-specific antigen as an example, exhibits remarkable efficiency by requiring only around 2 μL of sample volume. The decrease in sample amount required is facilitated through the application of nature inspired wettability patterning technique using PDMS-TiO2 nanocomposite coating, which incorporates distinct super-hydrophilic and super-hydrophobic regions on the electrode surface. This surface modification facilitates a highly sensitive antigen detection in minimal sample volumes. The sensor strip demonstrates a linear detection range spanning from 0.0001 to 100 ng mL-1, with a limit of detection of 0.0001 ng mL-1. Additionally, the sensor performance remains consistent across varying inclinations of the sensor strip, ensuring its applicability in the field of wearable sensing. Moreover, the incorporation of the PDMS-TiO2 coating enhances the electrochemical performance of the sensor by providing an extended surface area without introducing electrical interference. A wide range of electrochemical methods, such as cyclic voltammetry, linear sweep voltammetry, chronoamperometry, and electrochemical impedance spectroscopy, have been utilized. In support of these experimental findings, simulation studies have explored droplet spreading dynamics upon variation of the droplet size and electrode surface inclination. The developed sensor strip can serve as a promising candidate for practical point-of-care testing applications, expanding its applicability to a broad spectrum of immunosensing applications targeting diverse antigens.

Surveillance of the respiratory syncytial virus outside infancy: impact of testing methods, a retrospective observational study.

The European Medicines Agency has approved several vaccines to protect the elderly against respiratory syncytial virus (RSV) infections. However, differences in performance between antigen and PCR tests, especially in adults, can make monitoring RSV difficult. This study aims to assess the impact of the chosen diagnostic methods on the surveillance of RSV. RSV and influenza test results obtained from July 2022 to June 2023 in a consolidated clinical laboratory in Brussels, Belgium, were collected. These results included antigen tests, quadruplex PCR tests and viral cultures on respiratory samples. Epidemiological trends related to the age of patients and the diagnostic methods were analysed. Among 14 761 RSV tests, the overall number of positive tests for infants until 1 year of age peaked on 5 November 2022 (67 per 7 days) whereas it peaked on 22 December 2022 for adults (33 per 7 days). Positive antigen tests peaked on 7 November 2022 (56 per 7 days) whereas positive PCRs peaked on 19 December 2022 (36 per 7 days). Nevertheless, the positivity rate of RSV PCRs had peaked 1 month previously. Infants were mainly diagnosed through antigen testing, contrary to older patients. The influenza epidemic was probably the cause of the increased use of a quadruplex PCR, leading to a delayed increase in the absolute number of PCRs positive for RSV. This study shows that the use of different diagnostic methods could lead to an erroneous representation of RSV epidemiology in adults due to the lack of sensitivity of antigen detection. RSV surveillance in the elderly should rely rather on molecular methods.

Semi-quantitative graphene chemiresistor enzyme immunoassay for simple and sensitive antigen detection

Graphene is an ideal platform for sensing biomolecule charges due to its high electron/hole mobilities and 2D nature. However, when directly detecting biomolecules on graphene surfaces, it is important to consider their charge magnitude and the impact of Debye screening, which limits the detectable biomolecule types. Therefore, we propose a graphene chemiresistor enzyme immunoassay that allows for the detection of all protein types, regardless of charge or size. In this study, we immobilized antibodies on rigid glass surfaces rather than graphene surfaces to simplify the washing process and reduce noise caused by solution flow on graphene. As an example, we used influenza virus nucleoprotein as the antigen and urease as the enzyme. By measuring changes in graphene's electrical conductivity resulting from ammonium ions generated by enzymatic reactions, we confirmed antigen presence. The responsiveness of the graphene chemiresistor varied depending on antigen concentration, successfully detecting 100 pM antigens in this experiment. Our semi-quantitative method offers a simple and highly sensitive electrical detection of antigens.

Light-mediated protein functionalization of photoclickable hydrogel interface for selective cell capture and dot blotting assay

The construction of versatile functional hydrogel interfaces holds promising prospects in biosensing and bioengineering. Herein, we introduced a light-induced protein conjugation strategy for on-demand surface modification of hydrogel interface based on the photoclick cyclization between primary amine and o-nitrobenzyl alcohol. We achieved the on-demand protein conjugation by grafting the molecular plugin, 4-(hydroxymethyl)-3-nitrobenzoic acid (HNBA), onto the hydrogel surface, followed by the mask-aided photoclick reaction with the protein of interest. This method enables the creation of protein patterns on hydrogel interface with a lower limit of pattern width at ∼70 μm. With this method, we demonstrated the surface engineering of epidermal growth factor (EGF) on hydrogel interface for selective capture of EGF receptor-positive cancer cells with an efficiency over 80%. Moreover, we applied the mask-aided photoclick conjugation method for antigen capture and developed a photoclickable hydrogel interface-based dot blotting assay. Due to the high-efficient antigen capture of photoclick conjugation, the photoclickable hydrogel interface-based dot blotting assay shows improved sensitivity for antigen detection with a limit of detection as 0.065 ng. We believed that this light-induced protein conjugation method holds the potential as a robust strategy for the construction of bioactive hydrogel interfaces for various bio-related applications.

Diagnostic Overview of Foot-And-Mouth Disease Viruse (FMDV)

Foot-and-mouth disease (FMD) is one of the highly contagious diseases of domestic animals. Sensitive, specific, and quick detection of FMD virus antigens and antibodies is an essential corner stone diagnostic tool at each tier of control strategy. Continued development of molecular and genetic technologies combined with novel analytical techniques will benefit many areas of FMD research and understanding. Genome sequencing provides excellent insight into the evolutionary and epidemiological dynamics of such viruses. The advent of next generation sequencing (NGS) technologies enables evolutionary studies of the entire viral swarm. Sequencing technologies have advanced uses of viral genomic data; which helps to understand the global distribution and Trans-boundary movements of Foot-and-mouth virus disease. Also sequencing information can be used to understand antigenic change within virus strains; therefore, the aim of this review is to assess various diagnostic techniques including conventional and advanced techniques both from agent and host response sides. Among the agent side virus isolation, RT-PCR, antigen ELISA and immune-chromatography strips are commonly used diagnostic tests. Whereas from the host side: infrared thermo gram, antibody ELISA and Virus neutralization are some mentions. Owing to current technological advancement: DNA sequencing and microarray, expressing a luciferase reporter and quantitative proteomics were reviewed. Therefore, potential confirmatory diagnostic test should be in combination with each other.

Sensitive Lateral Flow Immunoassay Based on Specific Peptide and Superior Oxidase Mimics with a Universal Dual-Mode Significant Signal Amplification.

Rapid and sensitive antigen detection using a lateral flow immunoassay (LFIA) is crucial for diagnosing infectious diseases due to its simplicity, speed, and user-friendly features. However, it remains a critical issue to explore specific biorecognition elements and powerful signal amplification. In this study, taking SARS-CoV-2 as a proof of concept, a specific peptide, WFLNDSELIML, binding to the SARS-CoV-2 spike (S) antigen was identified by a nonamplified biopanning method, which exhibited high affinity to the target, with a dissociation constant of 9.29 ± 1.55 nM. Molecular docking analysis reveals that this peptide binds to the N-terminal domain of the SARS-CoV-2 S antigen. Then, using this peptide as a capture probe and angiotensin-converting enzyme 2 as a detection probe, a peptide-based lateral flow immunoassay (pLFIA) for the sensitive detection of the SARS-CoV-2 S antigen without any antibody was developed, for which a polydopamine nanosphere (PDA)@MnO2 nanocomposite with excellent oxidase-like activity was used as a colorimetric label, exhibiting dual-mode remarkable signal amplification of natural melanin and on-demand nanozyme catalytic enhancement. The PDA@MnO2-based pLFIA is capable of detecting the SARS-CoV-2 S antigen with a limit of detection of 8.01 pg/mL, which is 18.7 times lower than that of a conventional pLFIA tagged with gold nanoparticles. Additionally, the as-proposed PDA@MnO2-based pLFIA can detect up to 150 transduction units/mL SARS-CoV-2 pseudoviruses spiked in saliva samples. Given the outstanding analytical performance, the proposed PDA@MnO2-based pLFIA may offer a reliable option for the rapid diagnosis of SARS-CoV-2.

Recent Advances in Quantum Dot-Based Lateral Flow Immunoassays for the Rapid, Point-of-Care Diagnosis of COVID-19.

The COVID-19 pandemic has spurred demand for efficient and rapid diagnostic tools that can be deployed at point of care to quickly identify infected individuals. Existing detection methods are time consuming and they lack sensitivity. Point-of-care testing (POCT) has emerged as a promising alternative due to its user-friendliness, rapidity, and high specificity and sensitivity. Such tests can be conveniently conducted at the patient's bedside. Immunodiagnostic methods that offer the rapid identification of positive cases are urgently required. Quantum dots (QDs), known for their multimodal properties, have shown potential in terms of combating or inhibiting the COVID-19 virus. When coupled with specific antibodies, QDs enable the highly sensitive detection of viral antigens in patient samples. Conventional lateral flow immunoassays (LFAs) have been widely used for diagnostic testing due to their simplicity, low cost, and portability. However, they often lack the sensitivity required to accurately detect low viral loads. Quantum dot (QD)-based lateral flow immunoassays have emerged as a promising alternative, offering significant advancements in sensitivity and specificity. Moreover, the lateral flow immunoassay (LFIA) method, which fulfils POCT standards, has gained popularity in diagnosing COVID-19. This review focuses on recent advancements in QD-based LFIA for rapid POCT COVID-19 diagnosis. Strategies to enhance sensitivity using QDs are explored, and the underlying principles of LFIA are elucidated. The benefits of using the QD-based LFIA as a POCT method are highlighted, and its published performance in COVID-19 diagnostics is examined. Overall, the integration of quantum dots with LFIA holds immense promise in terms of revolutionizing COVID-19 detection, treatment, and prevention, offering a convenient and effective approach to combat the pandemic.

Cu-MOFs/GOx Bifunctional Probe-Based Synergistic Signal Amplification Strategy: Toward Highly Sensitive Closed Bipolar Electrochemiluminescence Immunoassay.

A closed bipolar electrochemiluminescence (BP-ECL) platform for sensitive prostate specific antigen (PSA) detection was proposed based on a novel synergistic signal amplification strategy. Specifically, glucose oxidase-loaded Cu-based metal-organic frameworks (Cu-MOFs/GOx) as bifunctional probes were bridged on the anodic interface with the target PSA as the intermediate unit. In virtue of the large loading capacity of Cu-MOFs, a large amount of a co-reactant, i.e., H2O2 in this L-012-based ECL system and gluconic acid were generated on the anodic pole in the presence of glucose. The generated gluconic acid could effectively degrade the Cu-MOFs to release Cu2+ which greatly accelerates the formation of highly active intermediates from co-reactant H2O2, boosting the ECL intensity. As for the cathodic pole, K3Fe(CN)6 with a lower reduction potential is used to reduce the driving voltage and speed up the reaction rate, further strengthening the ECL intensity. Thanks to the synergistic signal amplification effect at both two electrode poles of the BP-ECL system, highly sensitive detection of PSA was realized with a detection limit of 5.0 × 10-14 g/mL and a wide linear range of 1.0 × 10-13-1.0 × 10-7 g/mL. The strategy provides a novel way for signal amplification in the BP-ECL biosensing field.

PdPtRu trimetallic nanozymes and application to electrochemical immunosensor for sensitive SARS-COV-2 antigen detection

Herein, a ternary PdPtRu nanodendrite as novel trimetallic nanozyme was reported, which possessed excellent peroxidase-like activity as well as electro-catalytic activity on account of the synergistic effect between the three metals. Based on the excellent electro-catalytic activity of trimetallic PdPtRu nanozyme toward the reduction of H2O2, the trimetallic nanozyme was applied to construct a brief electrochemical immunosensor for SARS-COV-2 antigen detection. Concretely, trimetallic PdPtRu nanodendrite was used to modify electrode surface, which not only generated high reduction current of H2O2 for signal amplification, but also provided massive active sites for capture antibody (Ab1) immobilization to construct immunosensor. In the presence of target SARS-COV-2 antigen, SiO2 nanosphere labeled detection antibody (Ab2) composites were introduced on the electrode surface according sandwich immuno-reaction. Due to the inhibitory effect of SiO2 nanosphere on the current signal, the current signal was decreased with the increasing target SARS-COV-2 antigen concentration. As a result, the proposed electrochemical immunosensor presented sensitive detection of SARS-COV-2 antigen with linear range from 1.0 pg/mL to 1.0 μg/mL and limit of detection down to 51.74 fg/mL. The proposed immunosensor provide a brief but sensitive antigen detection tool for rapid diagnosis of COVID-19.

SERS/photothermal-based dual-modal lateral flow immunoassays for sensitive and simultaneous antigen detection of respiratory viral infections

Lateral flow immunoassay (LFIA) is one of the most common analytical platforms for point-of-care testing (POCT), which is capable of large-scale primary screening and home self-testing of infectious diseases. However, the sensitivity of conventional AuNPs-based LFIA is relatively low and more prone to false negatives. Herein, we report a novel LFIA based on gold-core-silver-shell bimetallic nanoparticles (Au4-ATP@Ag NPs) emitting Surface-enhanced Raman scatting (SERS) and Photothermal (PT) effect, named SERS/PT-based dual-modal LFIA (SERS/PT-dmLFIA), for the antigen detection of infectious diseases pathogens, which displayed an excellent performance. For influenza A virus (IAV), influenza B virus (IBV), and Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) N protein detection, the limit of detections (LoD) with Raman as signal were 31.25, 93.75, and 31.25 pg mL-1 respectively, and the LoDs with temperature difference (∆T) as signal were as low as 15.63, 187.5, and 15.63 pg mL-1 respectively, which were over 4-fold more sensitive than visual-based LFIA. The proposed SERS/PT-dmLFIA was used for detecting virus antigen in pharyngeal swabs and showed ideal coincidence rate of over 95% compared to the commercialized assays. In addition, we explored the development of multiplex SERS/PT-dmLFIA that can detect IAV, IBV, and SARS-CoV-2 antigens simultaneously without cross reactivity. Overall, the SERS/PT-dmLFIA for antigen detection not only exhibits high sensitivity, accuracy and specificity, but also have characteristics of rapidity and simplicity, which holds high potential for rapid diagnosis of infectious diseases in laboratory testing, mass screening, and home self-testing.

Local Sensing of Absolute Refractive Index During Protein‐Binding using Microlasers with Spectral Encoding

AbstractMultiplexed, specific, and sensitive detection of antigens is critical for the rapid and accurate diagnosis of disease and the informed development of personalized treatment plans. Here, it is shown that polymer microsphere lasers can be used as photonic sensors to monitor and quantify direct surface binding of biomolecules via changes in the refractive index. The unique spectral signature of each individual laser can be used to find their size and effective refractive index which adds a new encoding dimension when compared to conventional fluorescent beads. Antibody‐functionalized microlasers selectively detect protein binding, as demonstrated for Immunoglobulin G and C‐reactive protein, and have the ability to resolve different stages of the multilayer surface modification. Moreover, by continuously monitoring single lasers, the possibility of real‐time monitoring of binding dynamics between antigens in solution phase and the immobilized antibodies is demonstrated. For multiplexed detection, the microlasers are employed in a flow cytometer configuration, with fast spectral detection and identification of microlasers with and without antigen binding. It is envisioned that by combining microlasers with well‐established surface modification chemistries and flow geometries, the multiplexing ability of microbead immunoassays can be strongly increased while also opening avenues for single‐cell profiling within heterogeneous cell populations.

Disposal Immunosensor for Sensitive Electrochemical Detection of Prostate-Specific Antigen Based on Amino-Rich Nanochannels Array-Modified Patterned Indium Tin Oxide Electrode.

Sensitive detection of prostate-specific antigens (PSA) in serum is essential for the prevention and early treatment of prostate cancer. Simple and disposable electrochemical immunosensors are highly desirable for screening and mobile detection of PSAs in high-risk populations. Here, an electrochemical immunosensor was constructed based on amino-rich nanochannels array-modified patterned, inexpensive, and disposable indium tin oxide (ITO) electrodes, which can be employed for the sensitive detection of PSA. Using an amino-group-containing precursor, a vertically ordered mesoporous silica nanochannel film (VMSF) containing amino groups (NH2-VMSF) was rapidly grown on ITO. When NH2-VMSF contained template surfactant micelle (SM), the outer surface of NH2-VMSF was directionally modified by aldehyde groups, which enabled further covalent immobilization of the recognitive antibody to prepare the immuno-recognitive interface. Owing to the charge-based selective permeability, NH2-VMSF can electrostatically adsorb negatively charged redox probes in solution (Fe(CN)63-/4-). The electrochemical detection of PSA is realized based on the mechanism that the antigen-antibody complex can reduce the diffusion of redox probes in solution to the underlying electrode, leading to the decrease in electrochemical signal. The constructed immunosensor can achieve sensitive detection of PSA in the range from 10 pg/mL to 1 μg/mL with a limit of detection (LOD) of 8.1 pg/mL. Sensitive detection of PSA in human serum was also achieved. The proposed disposable immunosensor based on cheap electrode and nanochannel array is expected to provide a new idea for developing a universal immunosensing platform for sensitive detection of tumor markers.