scholarly journals Spectral image contrast-based flow digital nanoplasmon-metry for ultrasensitive antibody detection

2022 ◽  
Vol 20 (1) ◽  
Author(s):  
Sheng-Hann Wang ◽  
Chia-Wen Kuo ◽  
Shu-Cheng Lo ◽  
Wing Kiu Yeung ◽  
Ting-Wei Chang ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Artificial Saliva ◽  
Sample Volume ◽  
Image Contrast ◽  
Antibody Detection ◽  
Digital Analysis ◽  
High Concentration ◽  
Spectral Image ◽  
Image Brightness ◽  
Detection Range

Abstract Background Gold nanoparticles (AuNPs) have been widely used in local surface plasmon resonance (LSPR) immunoassays for biomolecule sensing, which is primarily based on two conventional methods: absorption spectra analysis and colorimetry. The low figure of merit (FoM) of the LSPR and high-concentration AuNP requirement restrict their limit of detection (LOD), which is approximately ng to μg mL−1 in antibody detection if there is no other signal or analyte amplification. Improvements in sensitivity have been slow in recent for a long time, and pushing the boundary of the current LOD is a great challenge of current LSPR immunoassays in biosensing. Results In this work, we developed spectral image contrast-based flow digital nanoplasmon-metry (Flow DiNM) to push the LOD boundary. Comparing the scattering image brightness of AuNPs in two neighboring wavelength bands near the LSPR peak, the peak shift signal is strongly amplified and quickly detected. Introducing digital analysis, the Flow DiNM provides an ultrahigh signal-to-noise ratio and has a lower sample volume requirement. Compared to the conventional analog LSPR immunoassay, Flow DiNM for anti-BSA detection in pure samples has an LOD as low as 1 pg mL−1 within only a 15-min detection time and 500 μL sample volume. Antibody assays against spike proteins of SARS-CoV-2 in artificial saliva that contained various proteins were also conducted to validate the detection of Flow DiNM in complicated samples. Flow DiNM shows significant discrimination in detection with an LOD of 10 pg mL−1 and a broad dynamic detection range of five orders of magnitude. Conclusion Together with the quick readout time and simple operation, this work clearly demonstrated the high sensitivity and selectivity of the developed Flow DiNM in rapid antibody detection. Spectral image contrast and digital analysis further provide a new generation of LSPR immunoassay with AuNPs. Graphical Abstract

scholarly journals Spectral Image Contrast Based Flow Digital Nanoplasmon-metry For Ultrasensitive Antibody Detection

2021 ◽  
Author(s):  
Sheng-Hann Wang ◽  
Chia-Wen Kuo ◽  
Shu-Cheng Lo ◽  
Wing Kiu Yeung ◽  
Ting-Wei Chang ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Artificial Saliva ◽  
Sample Volume ◽  
Image Contrast ◽  
Antibody Detection ◽  
High Signal ◽  
Digital Analysis ◽  
High Concentration ◽  
Spectral Image ◽  
Image Brightness

Abstract Background: Gold nanoparticles (AuNPs) have been widely used as local surface plasmon resonance (LSPR) immunoassay for biomolecule sensings. It is primarily based on two conventional methods - absorption spectra and colorimetric. While the low figure of merit (FoM) of the LSPR and high-concentration AuNPs requirement restrict their limit of detection (LOD), which is around ng to μg per mL in antibody detection if there is no other signal or analytes amplification. By now, the improvement of sensitivity has bogged down for years. It also reveals a great challenge of the current LSPR immunoassay in biosensing - pushing the boundary of the current LOD.Results: In this work, we developed a spectral image contrast-based flow digital nanoplasmon-metry(FDNM) to push the LOD boundary. Comparing the scattering image brightness of AuNPs in two neighboring wavelength bands near the LSPR peak, the signal of peak shift is extremely amplified and quickly detected. Introducing the digital analysis, the FDNM provides an ultra-high signal-to-noise ratio and less sample volume requirement. Compared to conventional analog LSPR immunoassay, FDNM has a LOD down to 1 pg mL-1 within only a 15-minute detection time and 500 μL sample volume. Antibody against spike proteins of SARS-CoV-2 in artificial saliva that contained various proteins was also conducted to validate the detection of FDNM in complicated samples. Of the detection, FDNM shows significant discrimination with a LOD of 10 pg mL-1 and a broad dynamic detecting range of five orders of magnitude. Conclusion: Together with the quick readout time and simple operation, this work outstandingly demonstrated the high sensitivity and selectivity of the developed FDNM in rapid antibody detection. The spectral image contrast and digital analysis further provide a new generation LSPR immunoassay of AuNPs.

scholarly journals An apta-aggregation based machine learning assay for rapid quantification of lysozyme through texture parameters

PLoS ONE ◽  
2021 ◽  
Vol 16 (3) ◽  
pp. e0248159
Author(s):  
Manoharan Sanjay ◽  
Kumar Gaurav ◽  
Maria Jesus Gonzalez-Pabon ◽  
Julio Fuchs ◽  
Susan R. Mikkelsen ◽  
...  
Keyword(s):  
Machine Learning ◽  
Limit Of Detection ◽  
Sample Volume ◽  
High Concentration ◽  
Experimental Procedure ◽  
Low Concentration ◽  
Operating Modes ◽  
Texture Parameters ◽  
Aggregation Property ◽  
Rapid Quantification

A novel assay technique that involves quantification of lysozyme (Lys) through machine learning is put forward here. This article reports the tendency of the well- documented Ellington group anti-Lys aptamer, to produce aggregates when exposed to Lys. This property of apta-aggregation has been exploited here to develop an assay that quantifies the Lys using texture and area parameters from a photograph of the elliptical aggregate mass through machine learning. Two assay sets were made for the experimental procedure: one with high Lys concentration between 25–100 mM and another with low concentration between 1–20 mM. The high concentration set had a sample volume of 10 μl while the low concentration set had a higher sample volume of 100 μl, in order to obtain the statistical texture values reliably from the aggregate mass. The platform exhibited an experimental limit of detection of 1 mM and a response time of less than 10 seconds. Further, two potential operating modes for the aptamer were hypothesized for this aggregation property and the more accurate mode among the two was ascertained through bioinformatics studies.

scholarly journals Validation of a Novel Diagnostic Approach Combining the VersaTREK™ System for Recovery and Real-Time PCR for the Identification of Mycobacterium chimaera in Water Samples

2021 ◽  
Vol 9 (5) ◽  
pp. 1031
Author(s):  
Roberto Zoccola ◽  
Alessia Di Blasio ◽  
Tiziana Bossotto ◽  
Angela Pontei ◽  
Maria Angelillo ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Limit Of Detection ◽  
Detection System ◽  
Bacterial Load ◽  
Microbial Control ◽  
Hospital Setting ◽  
Sample Volume ◽  
Analytical Sensitivity ◽  
Initial Water

Mycobacterium chimaera is an emerging pathogen associated with endocarditis and vasculitis following cardiac surgery. Although it can take up to 6–8 weeks to culture on selective solid media, culture-based detection remains the gold standard for diagnosis, so more rapid methods are urgently needed. For the present study, we processed environmental M. chimaera infected simulates at volumes defined in international guidelines. Each preparation underwent real-time PCR; inoculates were placed in a VersaTREK™ automated microbial detection system and onto selective Middlebrook 7H11 agar plates. The validation tests showed that real-time PCR detected DNA up to a concentration of 10 ng/µL. A comparison of the isolation tests showed that the PCR method detected DNA in a dilution of ×102 CFU/mL in the bacterial suspensions, whereas the limit of detection in the VersaTREK™ was <10 CFU/mL. Within less than 3 days, the VersaTREK™ detected an initial bacterial load of 100 CFU. The detection limit did not seem to be influenced by NaOH decontamination or the initial water sample volume; analytical sensitivity was 1.5 × 102 CFU/mL; positivity was determined in under 15 days. VersaTREK™ can expedite mycobacterial growth in a culture. When combined with PCR, it can increase the overall recovery of mycobacteria in environmental samples, making it potentially applicable for microbial control in the hospital setting and also in environments with low levels of contamination by viable mycobacteria.

scholarly journals A Compact Detection Platform Based on Gradient Guided-Mode Resonance for Colorimetric and Fluorescence Liquid Assay Detection

Sensors ◽  
2021 ◽  
Vol 21 (8) ◽  
pp. 2797
Author(s):  
Jing-Jhong Gao ◽  
Ching-Wei Chiu ◽  
Kuo-Hsing Wen ◽  
Cheng-Sheng Huang
Keyword(s):  
Limit Of Detection ◽  
Detection System ◽  
Fluorescence Emission ◽  
Assay Sensitivity ◽  
Detection Range ◽  
Current Form ◽  
Guided Mode ◽  
Spectral Detection ◽  
Guided Mode Resonance ◽  
Colorimetric Assays

This paper presents a compact spectral detection system for common fluorescent and colorimetric assays. This system includes a gradient grating period guided-mode resonance (GGP-GMR) filter and charge-coupled device. In its current form, the GGP-GMR filter, which has a size of less than 2.5 mm, can achieve a spectral detection range of 500–700 nm. Through the direct measurement of the fluorescence emission, the proposed system was demonstrated to detect both the peak wavelength and its corresponding intensity. One fluorescent assay (albumin) and two colorimetric assays (albumin and creatinine) were performed to demonstrate the practical application of the proposed system for quantifying common liquid assays. The results of our system exhibited suitable agreement with those of a commercial spectrometer in terms of the assay sensitivity and limit of detection (LOD). With the proposed system, the fluorescent albumin, colorimetric albumin, and colorimetric creatinine assays achieved LODs of 40.99 and 398 and 25.49 mg/L, respectively. For a wide selection of biomolecules in point-of-care applications, the spectral detection range achieved by the GGP-GMR filter can be further extended and the simple and compact optical path configuration can be integrated with a lab-on-a-chip system.

scholarly journals Demonstration of a Label-Free and Low-Cost Optical Cavity-Based Biosensor Using Streptavidin and C-Reactive Protein

Biosensors ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 4
Author(s):  
Donggee Rho ◽  
Seunghyun Kim
Keyword(s):  
Limit Of Detection ◽  
Point Of Care ◽  
Low Cost ◽  
Optical Cavity ◽  
Sample Volume ◽  
Small Sample ◽  
Label Free ◽  
C Reactive Protein ◽  
Reactive Protein ◽  
Cavity Structure

An optical cavity-based biosensor (OCB) has been developed for point-of-care (POC) applications. This label-free biosensor employs low-cost components and simple fabrication processes to lower the overall cost while achieving high sensitivity using a differential detection method. To experimentally demonstrate its limit of detection (LOD), we conducted biosensing experiments with streptavidin and C-reactive protein (CRP). The optical cavity structure was optimized further for better sensitivity and easier fluid control. We utilized the polymer swelling property to fine-tune the optical cavity width, which significantly improved the success rate to produce measurable samples. Four different concentrations of streptavidin were tested in triplicate, and the LOD of the OCB was determined to be 1.35 nM. The OCB also successfully detected three different concentrations of human CRP using biotinylated CRP antibody. The LOD for CRP detection was 377 pM. All measurements were done using a small sample volume of 15 µL within 30 min. By reducing the sensing area, improving the functionalization and passivation processes, and increasing the sample volume, the LOD of the OCB are estimated to be reduced further to the femto-molar range. Overall, the demonstrated capability of the OCB in the present work shows great potential to be used as a promising POC biosensor.

scholarly journals Ratiometric Fluorescent Biosensors for Glucose and Lactate Using an Oxygen-Sensing Membrane

Biosensors ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 208
Author(s):  
Hong Dinh Duong ◽  
Jong Il Rhee
Keyword(s):  
Limit Of Detection ◽  
Oxygen Sensing ◽  
High Sensitivity ◽  
Glucose Sensing ◽  
Oxidation Reactions ◽  
Lactate Oxidase ◽  
Lactate Oxidation ◽  
Detection Range ◽  
Quenching Effect ◽  
Sensing Membrane

In this study, ratiometric fluorescent glucose and lactate biosensors were developed using a ratiometric fluorescent oxygen-sensing membrane immobilized with glucose oxidase (GOD) or lactate oxidase (LOX). Herein, the ratiometric fluorescent oxygen-sensing membrane was fabricated with the ratio of two emission wavelengths of platinum meso-tetra (pentafluorophenyl) porphyrin (PtP) doped in polystyrene particles and coumarin 6 (C6) captured into silica particles. The operation mechanism of the sensing membranes was based on (i) the fluorescence quenching effect of the PtP dye by oxygen molecules, and (ii) the consumption of oxygen levels in the glucose or lactate oxidation reactions under the catalysis of GOD or LOX. The ratiometric fluorescent glucose-sensing membrane showed high sensitivity to glucose in the range of 0.1–2 mM, with a limit of detection (LOD) of 0.031 mM, whereas the ratiometric fluorescent lactate-sensing membrane showed the linear detection range of 0.1–0.8 mM, with an LOD of 0.06 mM. These sensing membranes also showed good selectivity, fast reversibility, and stability over long-term use. They were applied to detect glucose and lactate in artificial human serum, and they provided reliable measurement results.

scholarly journals Label-Free Electrochemical Biosensor Based on Au@MoS₂–PANI for Escherichia coli Detection

Chemosensors ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 49
Author(s):  
Pushap Raj ◽  
Man Hwan Oh ◽  
Kyudong Han ◽  
Tae Yoon Lee
Keyword(s):  
Escherichia Coli ◽  
Bacterial Infections ◽  
Limit Of Detection ◽  
Bacterial Pathogens ◽  
Cost Effective ◽  
Covalent Immobilization ◽  
Label Free ◽  
Self Assembled Monolayer ◽  
Detection Range ◽  
Linear Detection

Bacterial infections have become a significant challenge in terms of public health, the food industry, and the environment. Therefore, it is necessary to address these challenges by developing a rapid, cost-effective, and easy-to-use biosensor for early diagnosis of bacterial pathogens. Herein, we developed a simple, label-free, and highly sensitive immunosensor based on electrochemical detection using the Au@MoS₂–PANI nanocomposite. The conductivity of the glassy carbon electrode is greatly enhanced using the Au@MoS₂–PANI nanocomposite and a self-assembled monolayer of mercaptopropionic acid on the gold nanoparticle surface was employed for the covalent immobilization of antibodies to minimize the nonspecific adsorption of bacterial pathogens on the electrode surface. The biosensor established a high selectivity and sensitivity with a low limit of detection of 10 CFU/mL, and detected Escherichia coli within 30 min. Moreover, the developed biosensor demonstrated a good linear detection range, practical utility in urine samples, and electrode regenerative studies.

scholarly journals Gold-nanourchin complexed silicon dioxide-probe on gap-fingered interdigitated electrode surface for Parkinson’s Disease determination by current–volt measurement

2021 ◽  
Vol 11 ◽  
pp. 184798042098735
Author(s):  
Xiaohong Li ◽  
Wei Shi ◽  
Wenyan Zhang ◽  
Weiyao Chen ◽  
Dan Cao ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Silicon Dioxide ◽  
Limit Of Detection ◽  
Detection System ◽  
Interdigitated Electrode ◽  
Limit Of Quantification ◽  
Neurofilament Light Chain ◽  
Neurofilament Light ◽  
Detection Range

Parkinson’s disease (PD) is a nervous disorder, affects physical movement, and leads to difficulty in balancing, walking, and coordination. A novel sensor is mandatory to determine PD and monitor the progress of the treatment. Neurofilament light chain (NfL) has been recognized as a good biomarker for PD and also helps to distinguish between PD and atypical PD syndromes. Immunosensor was generated by current–volt measurement on gap-fingered interdigitated electrode with silicon dioxide surface to determine NfL level. To enhance the detection, anti-NfL antibody was complexed with gold-nanourchin and immobilized on the sensing electrode. The current–volt response was gradually increased at the linear detection range from 100 fM to 1 nM. Limit of detection and sensitivity were 100 fM with the signal-to-noise ratio at n = 3 on a linear curve ( y = 0.081 x + 1.593; R 2 = 0.9983). Limit of quantification falls at 1 pM and high performance of the sensor was demonstrated by discriminating against other neurogenerative disease markers, in addition, it was reproducible even in serum-spiked samples. This method of detection system aids to measure the level of NfL and leads to determine the condition with PD.

scholarly journals Streptavidin-conjugated gold nanoclusters as ultrasensitive fluorescent sensors for early diagnosis of HIV infection

2018 ◽  
Vol 4 (11) ◽  
pp. eaar6280 ◽  
Author(s):  
Aditya Dileep Kurdekar ◽  
L. A. Avinash Chunduri ◽  
C. Sai Manohar ◽  
Mohan Kumar Haleyurgirisetty ◽  
Indira K. Hewlett ◽  
...  
Keyword(s):  
Hiv Infection ◽  
Limit Of Detection ◽  
Gold Nanoclusters ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dependent Manner ◽  
Metal Nanoclusters ◽  
Detection Range ◽  
In Silico Studies ◽  
P24 Antigen ◽  
Hiv 1

We have engineered streptavidin-labeled fluorescent gold nanoclusters to develop a gold nanocluster immunoassay (GNCIA) for the early and sensitive detection of HIV infection. We performed computational simulations on the mechanism of interaction between the nanoclusters and the streptavidin protein via in silico studies and showed that gold nanoclusters enhance the binding to the protein, by enhancing interaction between the Au atoms and the specific active site residues, compared to other metal nanoclusters. We also evaluated the role of glutathione conjugation in binding to gold nanoclusters with streptavidin. As proof of concept, GNCIA achieved a sensitivity limit of detection of HIV-1 p24 antigen in clinical specimens of 5 pg/ml, with a detection range up to1000 pg/ml in a linear dose-dependent manner. GNCIA demonstrated a threefold higher sensitivity and specificity compared to enzyme-linked immunosorbent assay for the detection of HIV p24 antigen. The specificity of the immunoassay was 100% when tested with plasma samples negative for HIV-1 p24 antigen and positive for viruses such as hepatitis B virus, hepatitis C virus, and dengue. GNCIA could be developed into a universal labeling technology using the relevant capture and detector antibodies for the specific detection of antigens of various pathogens in the future.

scholarly journals Butyl Benzyl Phthalate in Urban Sewage by Magnetic-Based Immunoassay: Environmental Levels and Risk Assessment

Biosensors ◽  
2022 ◽  
Vol 12 (1) ◽  
pp. 45
Author(s):  
Xia Hong ◽  
Yin Cui ◽  
Ming Li ◽  
Yifan Xia ◽  
Daolin Du ◽  
...  
Keyword(s):  
Risk Assessment ◽  
Drinking Water ◽  
Limit Of Detection ◽  
Environmental Water ◽  
Carcinogenic Risk ◽  
Essential Information ◽  
Detection Range ◽  
Urban Sewage ◽  
Butyl Benzyl Phthalate ◽  
Positive Rate

A magnetic-based immunoassay (MBI) combined with biotin-streptavidin amplification was proposed for butyl benzyl phthalate (BBP) investigation and risk assessment. The values of LOD (limit of detection, IC10) and IC50 were 0.57 ng/mL and 119.61 ng/mL, with a detection range of 0.57–24977.71 ng/mL for MBI. The specificity, accuracy and precision are well demonstrated. A total of 36 environmental water samples of urban sewage from Zhenjiang, China, were collected and assessed for BBP contamination. The results show that BBP-positive levels ranged from 2.47 to 89.21 ng/mL, with a positive rate of 77.8%. The health effects of BBP in the urban sewage were within a controllable range, and the ambient severity for health (ASI) was below 1.49. The highest value of AS for ecology (ASII) was 7.43, which indicates a potential harm to ecology. The entropy value of risk quotient was below 100, the highest being 59.47, which poses a low risk to the environment and ecology, indicating that there is a need to strengthen BBP controls. The non-carcinogenic risk of BBP exposure from drinking water was higher for females than that for males, and the non-carcinogenic risk from drinking-water and bathing pathways was negligible. This study could provide an alternative method for detecting BBP and essential information for controlling BBP contamination.

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