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Turbidimetric inhibition immunoassay revisited to enhance its sensitivity via an optofluidic laser
摘要： Turbidimetric inhibition immunoassay (TIIA) is a classic immunodiagnostic method that has been extensively used for biomarker detection. However, the low sensitivity of this technique hinders its applications in the early diagnosis of diseases. Here, a new concept, optofluidic laser TIIA (OFL-TIIA), is proposed and demonstrated for sensitive protein detection. In contrast to the immunoreaction in traditional TIIA, in which the single-pass laser loss is detected, the immunoreaction in the OFL-TIIA method takes place in a laser cavity, which considerably increases the loss induced by antigen-antibody complexes (AACs) via the amplification effect of the laser. A commercial IgG TIIA kit was selected as a demonstrative model to characterize the performance of OFL-TIIA. A wide dynamic range of five orders of magnitude with an exceptional limit of detection (LOD) (1.8×10-10 g/L) was achieved. OFL-TIIA is a fast, sensitive, and low-cost immunoassay with a simple homogeneous and wash-free process and low-volume sample consumption, thus providing a new detection platform for disease diagnostics.
关键词： Biomarker detection,Optofluidic laser,Turbidimetric inhibition immunoassay,Antigen-antibody complexes,Laser dye
更新于2025-11-25 10:30:42
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Silanized quantum dots as labels in lateral flow test strips for C-reactive protein
摘要： The paper describes the first use of silanized semiconductor core-shell quantum dots as fluorescent labels for macromolecule, C-reactive protein determination in blood plasma. The controlled synthesis of CdSe cores, with successive shells of CdS, CdZnS, ZnS and coating with transparent, stable, and inert silica shell, provides quantum dots with a narrow emission band, high quantum yield, and prolonged signal stability. Finally, the quantum dots were conjugated with specific antibodies via carboxylic groups on the silica surface. The method was further used for the immunochromatographic assay of C-reactive protein, a diagnostically important inflammatory biomarker. Assays with both the fluorescent QDs and a widely used colloidal gold label were developed in parallel and compared. The silanized quantum dots provide a more sensitive assay with a detection limit of 1 ng/mL for C-reactive protein in standard solutions, whereas the common assay has a detection limit of 10 ng/mL. The possibility of quantitative evaluation of analyte content by a portable device was demonstrated; the accuracy of the measurements was in the range of 5%–10%. The tests were used to determine C-reactive proteins in human plasma samples. The selected optimized protocol for these samples is based on a 4-fold dilution. The final working range of the assay, 4–1,200 ng/mL, covers practically all important interval of C-reactive protein values for the characterization of acute, chronic, and local inflammatory processes. Due to their high physical stability and inertness as well as intense, stable, and reproducible fluorescence, silanized quantum dots may be applied for high-sensitive assays for different analytes.
关键词： C-reactive protein,Quantum dots,silanization,lateral flow immunoassay
更新于2025-11-19 16:46:39
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Development of a fluorescence immunoassay for highly sensitive detection of amantadine using the nanoassembly of carbon dots and MnO2 nanosheets as the signal probe
摘要： Fluorescence immunoassays are rapid, convenient and cost-effective for the sensitive quantitation of chemical contaminants in foodstuff. In this study, a competitive fluorescence ELISA was developed for the sensitive detection of amantadine (AMD) based on the alkaline phosphatase (ALP)-triggered fluorescence "turn-on" signals. As a fluorescence substrate, carbon dots (CDs) were adsorbed onto the surface of the MnO2 nanosheets (NSs) and formed a nanoassembly of p-CDs@MnO2 NSs which results in the fluorescence quench of CDs. The ALP labelled on antibody could catalyze the hydrolysis of the 2-phospho-L-ascorbic acid into ascorbic acid. The latter could then reduce and decompose the MnO2 NSs, which was accompanied by the release of CDs from the surface of MnO2 NSs and led to the fluorescence recovery of CDs. The change of the fluorescence intensity is related to the concentration of AMD in solution and thus could be applied to detect AMD in an ALP-based ELISA system. The fluorescent ELISA showed a linear detection for AMD in the range of 0.048 ng mL?1 to 1.1 ng mL?1 with a detection limit (LOD) of 0.035 ng mL?1. The novel fluorescent ELISA shows potential for the highly sensitive detection of AMD and other analytes in food analysis.
关键词： manganese dioxide nanosheets,carbon dots,fluorescent immunoassay,amantadine
更新于2025-11-14 17:04:02
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A smartphone-based system for fluorescence polarization assays
摘要： This paper demonstrates the use of a smartphone-based sensor for fluorescence polarization (FP) analysis of biomolecules. The FP detection can rapidly sense ligand-analyte bindings by measuring molecule mobility, and thus, FP-based assays have been widely used for rapid diagnostics in clinics. Here, we implemented the FP detection apparatus using a 3D-printed compact holder and the built-in camera of a smartphone. The system offers accurate measurements of the degree of polarization by simultaneously detecting the fluorescence intensities parallel and perpendicular to the polarization of the excitation. The fluorescence signal of the sample is excited by a laser or light-emitting diode and separated by a polarization beam cube depending on the polarization. Parallel and perpendicular polarized emissions are projected onto two different regions of the sensor chip in the smartphone camera. A custom software app was developed to count the average intensity in the areas of interest and compute the degree of polarization. We validated the system by measuring the polarization of dye molecules dissolved in solutions with different viscosities. As an example of biomolecule sensing, a competitive FP immunoassay of Prostaglandin E2 was demonstrated using the developed system and exhibited the limit of detection of 1.57 ng/mL. The smartphone-based FP assay platform can also be implemented for the detection of toxins, disease biomarkers, and pathogens in resource-limited settings.
关键词： immunoassay,mobile sensor,fluorescence polarization assay,smartphone
更新于2025-09-23 15:23:52
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An infrared IgG immunoassay based on the use of a nanocomposite consisting of silica coated Fe3O4 superparticles
摘要： A reliable, rapid and ultrasensitive immunoassay is described for determination of immunoglobulin G (IgG). It is making use of biofunctional magnetite (Fe3O4) superparticles coated with SiO2 and serving as an infrared (IR) probe. The unique IR fingerprint signals originating from the transverse and longitudinal phonon modes, respectively, of the asymmetric stretching of the Si–O–Si bridges display a satisfactory resistance to optical interference from the environment. The adoption of Fe3O4 superparticles instead of Fe3O4 nanoparticles as the magnetic core warrants a controllable structure and a strong magnetic response. This facilitates the efficient purification of the probes and the alleviation of the interfacial resistance between the liquid-solid interfaces by using a magnet. The gold-coated substrate was used to immobilize goat-anti-human IgG. The analyte (human IgG) was incubated with the IR probes, and then captured by the substrate immobilized antibody with the assistance of an external magnetic field. The integral area of the IR absorption band between 1250 cm?1 – 900 cm?1 was chosen for quantitative assay. The limit of detection is 95 fM, which is two orders of magnitude better than that without the magnetic field. The assay time was shortened from 2 h to 1 min. High selectivity, specificity, and long-term stability of the immunoassay were achieved. The performance of the assay when analyzing blood samples confirmed the practicability of the method.
关键词： Molecular vibration,IR spectroscopy,Core-shell,Sandwich immunoassay,Self-assembly,Protein,Blood,Superparamagnetism,Magnetic beads
更新于2025-09-23 15:23:52
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Smartphone-Based Fluorescent Lateral Flow Immunoassay Platform for Highly Sensitive Point-of-Care Detection of Zika Virus Nonstructural Protein 1
摘要： Simple, inexpensive, and rapid diagnostic tests in low-resource settings with limited laboratory equipment and technical expertise are instrumental in reducing morbidity and mortality from epidemic infectious diseases. We developed a smartphone-based fluorescent lateral flow immunoassay (LFIA) platform for the highly sensitive point-of-care detection of Zika virus nonstructural protein 1 (ZIKV NS1). An attachment was designed and 3D-printed to integrate the smartphone with external optical and electrical components, enabling the miniaturization of the instrument and reduction in cost and complexity. Quantum dot microspheres were utilized as probes in fluorescent LFIA because of their extremely bright fluorescence signal. This approach can achieve quantitative point-of-care detection of ZIKV NS1 within 20 min. Limits of detection (LODs) in buffer and serum were 0.045 and 0.15 ng mL-1, respectively. Despite the high structural similarity, a high-level Dengue virus NS1 as interferent showed limited cross-reactivity. Furthermore, this assay was successfully applied to detecte ZIKV NS1 and virions spiked in complex biological samples, indicating its practical application capability. Given its low cost, compact size, and excellent analytical performance, the proposed smartphone-based fluorescent LFIA platform holds considerable potential in rapid and accurate point-of-care detection of ZIKV NS1 and provides new insight into the design and application of molecular diagnostic methods in low-resource settings.
关键词： Quantum dot microsphere,Smartphone,Lateral flow immunoassay,Zika virus nonstructural protein 1,Point-of-care
更新于2025-09-23 15:23:52
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Ultrasensitive detection of avian influenza A (H7N9) virus using surface-enhanced Raman scattering-based lateral flow immunoassay strips
摘要： The development of biosensors that are portable, low-cost, and quantitative has long been sought for rapid, on-site, and timely detection of avian influenza virus (AIV). In this study, an antibody-based Raman lateral flow immunoassay strip was developed to detect AIV H7N9. This LFIA strip used a novel core-shell structure material, AuAg4(cid:3)ATP@AgNPs, as a Raman probe. An antibody specific for AIV and goat anti-mouse IgG antibody were immobilized on a nitrocellulose membrane as the test and control lines, respectively. Accumulation of antibody-virus-antibody-Raman probe complex at the test line could be visualized by the naked eye, and the Raman signal could be quantified using a portable Raman instrument. The testing process for the SERS-based LFIA strips could be completed in 20 min, which avoided the time-cost of current methods for AIV analysis. In our SERS-based biosensor, we estimated the limit of detection (LOD) for H7N9 to be 0.0018 HAU. This value is approximately three orders of magnitude more sensitive than the corresponding HA assays. When testing real sample, the results of the strip test were in accordance with those from real-time PCR testing. In conclusion, the SERS-based LFIA strip proposed in this study shows tremendous potential to detect targets quickly and sensitively using an elegantly simple method.
关键词： AuAg4(cid:3)ATP@AgNPs,Surface-enhanced Raman scattering,Avian influenza virus,Lateral flow immunoassay strips
更新于2025-09-23 15:23:52
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[IEEE 2018 IEEE Life Sciences Conference (LSC) - Montreal, QC, Canada (2018.10.28-2018.10.30)] 2018 IEEE Life Sciences Conference (LSC) - Assay Development and Storage for Fluorescence-Based Lateral Flow Immunoassay
摘要： Point-of-care medical diagnostics can provide efficient, cost-effective medical care, and have the potential to fundamentally change our current approach to global health. There have been substantial efforts in developing lateral flow assays for serologic testing, but most of the existing approaches have limited portability, are expensive, and offer limited analytical sensitivity. In this paper, we demonstrated an assay for the detection of antibodies in plasma to Epstein-Barr Nuclear Antigen-1 (EBNA-1) protein and optimization of the assay including washing and blocking conditions. We also investigated the effect of the storage on the assay strips. Using our optimized conditions, we were able to detect anti-Human Papilloma Virus (HPV)-16 E7 antibodies after three weeks of storage. Our goal is to adapt this system to detect HPV biomarkers for cervical cancers in low and middle-income countries.
关键词： storage,immunoassay,sensitivity,Lateral flow assay,fluorescence,point-of-care
更新于2025-09-23 15:22:29
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Switchable fluorescence immunoassay using gold nanoclusters anchored cobalt oxyhydroxide composite for sensitive detection of imidacloprid
摘要： The fabrication of a convenient fluorescence-based immunoassay (FIA) for specific antigen attract increasing concern but remains a considerable challenge, especially the laborious synthesis of nanomaterials-antibody conjugates. Herein, we circumvent this drawback by introducing a specific fluorescence signal response procedure to commercially available alkaline phosphatase (ALP)-labeled conventional immunoassay for imidacloprid detection. In this FIA protocol, gold nanoclusters (AuNCs) can anchor on the surface of two-dimensional cobalt oxyhydroxide (CoOOH) nanoflakes to form nanocomposite, resulting in remarkable decrease of fluorescence intensity. The quenching effects can be effectively reversed by introducing ascorbic acid that can trigger the decomposition of CoOOH nanoflakes. Notably, the corresponding fluorescence response induced by ascorbic acid was related to ALP activities labeled on antibody. After competitive immunoreaction, the ALP-labeled antibodies were bounded to immobilized antigen, which can regulate the fluorescence change. Utilizing fluorescence switching of system, the 50% inhibition concentration (IC50) value of FIA toward imidacloprid was obtained at 1.3 ng mL?1 which was 60-fold-of-magnitude more sensitive than that of conventional ELISA (86.4 ng mL?1). This FIA protocol not only develops new prospects for pesticide detection, but also opens up potent strategy of fluorogenic immunoassay.
关键词： Immunoassay,Composite,Alkaline phosphatase,Fluorescence,Imidacloprid
更新于2025-09-23 15:22:29
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Enlargement of Gold Nanoparticles for Sensitive Immunochromatographic Diagnostics of Potato Brown Rot
摘要： Lateral ?ow immunoassay (LFIA) is a convenient tool for rapid ?eld-based control of various bacterial targets. However, for many applications, the detection limits obtained by LFIA are not suf?cient. In this paper, we propose enlarging gold nanoparticles’ (GNPs) size to develop a sensitive lateral ?ow immunoassay to detect Ralstonia solanacearum. This bacterium is a quarantine organism that causes potato brown rot. We fabricated lateral ?ow test strips using gold nanoparticles (17.4 ± 1.0 nm) as a label and their conjugates with antibodies speci?c to R. solanacearum. We proposed a signal enhancement in the test strips’ test zone due to the tetrachloroauric (III) anion reduction on the GNP surface, and the increase in size of the gold nanoparticles on the test strips was approximately up to 100 nm, as con?rmed by scanning electron microscopy. Overall, the gold enhancement approach decreased the detection limit of R. solanacearum by 33 times, to as low as 3 × 104 cells·mL–1 in the potato tuber extract. The achieved detection limit allows the diagnosis of latent infection in potato tubers. The developed approach based on gold enhancement does not complicate analyses and requires only 3 min. The developed assay together with the sample preparation and gold enlargement requires 15 min. Thus, the developed approach is promising for the development of lateral ?ow test strips and their subsequent introduction into diagnostic practice.
关键词： gold nanoparticles,immunochromatographic diagnostics,potato brown rot,gold particle growth,increase of sensitivity,lateral ?ow immunoassay,test strips,Ralstonia solanacearum
更新于2025-09-23 15:22:29