scholarly journals A plug-and-play platform of ratiometric bioluminescent sensors for homogeneous immunoassays

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yan Ni ◽  
Bas J. H. M. Rosier ◽  
Eva A. van Aalen ◽  
Eva T. L. Hanckmann ◽  
Lieuwe Biewenga ◽  
...  
Keyword(s):  
Point Of Care ◽  
Low Cost ◽  
Spike Protein ◽  
Protein G ◽  
Therapeutic Antibodies ◽  
Protein Biomarkers ◽  
Plug And Play ◽  
External Calibration ◽  
Quantitative Measurements ◽  
Clinical Laboratories

AbstractHeterogeneous immunoassays such as ELISA have become indispensable in modern bioanalysis, yet translation into point-of-care assays is hindered by their dependence on external calibration and multiple washing and incubation steps. Here, we introduce RAPPID (Ratiometric Plug-and-Play Immunodiagnostics), a mix-and-measure homogeneous immunoassay platform that combines highly specific antibody-based detection with a ratiometric bioluminescent readout. The concept entails analyte-induced complementation of split NanoLuc luciferase fragments, photoconjugated to an antibody sandwich pair via protein G adapters. Introduction of a calibrator luciferase provides a robust ratiometric signal that allows direct in-sample calibration and quantitative measurements in complex media such as blood plasma. We developed RAPPID sensors that allow low-picomolar detection of several protein biomarkers, anti-drug antibodies, therapeutic antibodies, and both SARS-CoV-2 spike protein and anti-SARS-CoV-2 antibodies. With its easy-to-implement standardized workflow, RAPPID provides an attractive, fast, and low-cost alternative to traditional immunoassays, in an academic setting, in clinical laboratories, and for point-of-care applications.

scholarly journals RAPPID: a platform of ratiometric bioluminescent sensors for homogeneous immunoassays

2020 ◽  
Author(s):  
Yan Ni ◽  
Bas J.H.M. Rosier ◽  
Eva A. van Aalen ◽  
Eva T.L. Hanckmann ◽  
Lieuwe Biewenga ◽  
...  
Keyword(s):  
Target Recognition ◽  
Point Of Care ◽  
Low Cost ◽  
Digital Camera ◽  
Spike Protein ◽  
Therapeutic Antibodies ◽  
Protein Biomarkers ◽  
External Calibration ◽  
Quantitative Measurements ◽  
Clinical Laboratories

AbstractHeterogeneous immunoassays such as ELISA have become indispensable in modern bioanalysis, yet translation into easy-to-use point-of-care assays is hindered by their dependence on external calibration and multiple washing and incubation steps. Here, we introduce RAPPID (Ratiometric Plug-and-Play Immunodiagnostics), a “mix-and-measure” homogeneous immunoassay platform that combines highly specific antibody-based detection with a ratiometric bioluminescent readout that can be detected using a basic digital camera. The concept entails analyte-induced complementation of split NanoLuc luciferase fragments, photoconjugated to an antibody sandwich pair via protein G adapters. We also introduce the use of a calibrator luciferase that provides a robust ratiometric signal, allowing direct in-sample calibration and quantitative measurements in complex media such as blood plasma. We developed RAPPID sensors that allow low-picomolar detection of several protein biomarkers, anti-drug antibodies, therapeutic antibodies, and both SARS-CoV-2 spike protein and anti-SARS-CoV-2 antibodies. RAPPID combines ratiometric bioluminescent detection with antibody-based target recognition into an easy-to-implement standardized workflow, and therefore represents an attractive, fast, and low-cost alternative to traditional immunoassays, both in an academic setting and in clinical laboratories for point-of-care applications.

scholarly journals A rotationally focused flow (RFF) microfluidic biosensor by density difference for early-stage detectable diagnosis

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Noori Kim ◽  
Kyungsup Han ◽  
Pei-Chen Su ◽  
Insup Kim ◽  
Yong-Jin Yoon
Keyword(s):  
Early Detection ◽  
Optical Sensors ◽  
Early Stage ◽  
Point Of Care ◽  
Low Cost ◽  
Ease Of Use ◽  
Wavelength Shift ◽  
Label Free ◽  
Protein Biomarkers ◽  
Microfluidic Biosensor

AbstractLabel-free optical biosensors have received tremendous attention in point-of-care testing, especially in the emerging pandemic, COVID-19, since they advance toward early-detection, rapid, real-time, ease-of-use, and low-cost paradigms. Protein biomarkers testings require less sample modification process compared to nucleic-acid biomarkers’. However, challenges always are in detecting low-concentration for early-stage diagnosis. Here we present a Rotationally Focused Flow (RFF) method to enhance sensitivity(wavelength shift) of label-free optical sensors by increasing the detection probability of protein-based molecules. The RFF is structured by adding a less-dense fluid to focus the target-fluid in a T-shaped microchannel. It is integrated with label-free silicon microring resonators interacting with biotin-streptavidin. The suggested mechanism has demonstrated 0.19 fM concentration detection along with a significant magnitudes sensitivity enhancement compared to single flow methods. Verified by both CFD simulations and fluorescent flow-experiments, this study provides a promising proof-of-concept platform for next-generation lab-on-a-chip bioanalytics such as ultrafast and early-detection of COVID-19.

scholarly journals The SARS-COV-2 Spike Protein Binds Sialic Acids, and Enables Rapid Detection in a Lateral Flow Point of Care Diagnostic Device

2020 ◽  
Author(s):  
Alexander N. Baker ◽  
Sarah-Jane Richards ◽  
Collette S. Guy ◽  
Thomas R. Congdon ◽  
Muhammad Hasan ◽  
...  
Keyword(s):  
Point Of Care ◽  
Low Cost ◽  
Lateral Flow ◽  
Function Optimization ◽  
Spike Protein ◽  
Spike Glycoprotein ◽  
Binding Function ◽  
Point Of Care Diagnostics ◽  
Novel Coronavirus ◽  
Flow Detection

<div> <div> <div> <p>There is an urgent need to understand the behavior of novel coronavirus (SARS-COV-2), which is the causative agent of COVID-19, and to develop point-of-care diagnostics. Here, a glyconanoparticle platform is used to discover that N-acetyl neuraminic acid has high affinity towards the SARS-COV-2 spike glycoprotein, demonstrating its glycan-binding function. Optimization of the particle size and coating enabled detection of the spike glycoprotein in lateral flow and showed selectivity over the SARS-COV-1 spike protein. Using a viral particle mimic, paper-based lateral flow detection was demonstrated in under 30 minutes showing the potential of this system as a low-cost detection platform. </p> </div> </div> </div>

scholarly journals 3D-Printed Immunosensor Arrays for Cancer Diagnostics

Sensors ◽  
2020 ◽  
Vol 20 (16) ◽  
pp. 4514 ◽  
Author(s):  
Mohamed Sharafeldin ◽  
Karteek Kadimisetty ◽  
Ketki S. Bhalerao ◽  
Tianqi Chen ◽  
James F. Rusling
Keyword(s):  
3D Printing ◽  
Early Stage ◽  
Point Of Care ◽  
Low Cost ◽  
Cancer Diagnostics ◽  
Diagnostic Tools ◽  
Protein Biomarkers ◽  
Protein Biomarker ◽  
Subtractive Manufacturing ◽  
3D Printed

Detecting cancer at an early stage of disease progression promises better treatment outcomes and longer lifespans for cancer survivors. Research has been directed towards the development of accessible and highly sensitive cancer diagnostic tools, many of which rely on protein biomarkers and biomarker panels which are overexpressed in body fluids and associated with different types of cancer. Protein biomarker detection for point-of-care (POC) use requires the development of sensitive, noninvasive liquid biopsy cancer diagnostics that overcome the limitations and low sensitivities associated with current dependence upon imaging and invasive biopsies. Among many endeavors to produce user-friendly, semi-automated, and sensitive protein biomarker sensors, 3D printing is rapidly becoming an important contemporary tool for achieving these goals. Supported by the widely available selection of affordable desktop 3D printers and diverse printing options, 3D printing is becoming a standard tool for developing low-cost immunosensors that can also be used to make final commercial products. In the last few years, 3D printing platforms have been used to produce complex sensor devices with high resolution, tailored towards researchers’ and clinicians’ needs and limited only by their imagination. Unlike traditional subtractive manufacturing, 3D printing, also known as additive manufacturing, has drastically reduced the time of sensor and sensor array development while offering excellent sensitivity at a fraction of the cost of conventional technologies such as photolithography. In this review, we offer a comprehensive description of 3D printing techniques commonly used to develop immunosensors, arrays, and microfluidic arrays. In addition, recent applications utilizing 3D printing in immunosensors integrated with different signal transduction strategies are described. These applications include electrochemical, chemiluminescent (CL), and electrochemiluminescent (ECL) 3D-printed immunosensors. Finally, we discuss current challenges and limitations associated with available 3D printing technology and future directions of this field.

scholarly journals The SARS-COV-2 Spike Protein Binds Sialic Acids, and Enables Rapid Detection in a Lateral Flow Point of Care Diagnostic Device

2020 ◽  
Author(s):  
Alexander N. Baker ◽  
Sarah-Jane Richards ◽  
Collette S. Guy ◽  
Thomas R. Congdon ◽  
Muhammad Hasan ◽  
...  
Keyword(s):  
Point Of Care ◽  
Low Cost ◽  
Lateral Flow ◽  
Function Optimization ◽  
Spike Protein ◽  
Spike Glycoprotein ◽  
Binding Function ◽  
Point Of Care Diagnostics ◽  
Novel Coronavirus ◽  
Flow Detection

<div> <div> <div> <p>There is an urgent need to understand the behavior of novel coronavirus (SARS-COV-2), which is the causative agent of COVID-19, and to develop point-of-care diagnostics. Here, a glyconanoparticle platform is used to discover that N-acetyl neuraminic acid has high affinity towards the SARS-COV-2 spike glycoprotein, demonstrating its glycan-binding function. Optimization of the particle size and coating enabled detection of the spike glycoprotein in lateral flow and showed selectivity over the SARS-COV-1 spike protein. Using a viral particle mimic, paper-based lateral flow detection was demonstrated in under 30 minutes showing the potential of this system as a low-cost detection platform. </p> </div> </div> </div>

A Node Localization Algorithm Used for POCT Microfluidic Detection

2013 ◽  
Vol 846-847 ◽  
pp. 684-687
Author(s):  
Chao Lin
Keyword(s):  
Point Of Care ◽  
Low Cost ◽  
Location Information ◽  
Localization Algorithm ◽  
Point Of Care Testing ◽  
Node Localization ◽  
Plug And Play ◽  
Microfluidic Technology

This paper intends to study in the POCT (point-of-care-testing) combined with microfluidic technology as the breakthrough point. A localization algorithm is used for the ultimate miniaturization, portable, low cost, plug-and-play type PERS detecting instrument, which consist of telemedicine perception layer, and then collected data and location information more effectively transmitted to the server to deal with and feedback.

scholarly journals Validation of a point-of-care rapid diagnostic test for hepatitis C for use in resource-limited settings

2019 ◽  
Vol 11 (4) ◽  
pp. 314-315
Author(s):  
James S Leathers ◽  
Maria Belen Pisano ◽  
Viviana Re ◽  
Gertine van Oord ◽  
Amir Sultan ◽  
...  
Keyword(s):  
Point Of Care ◽  
Low Cost ◽  
Rapid Diagnosis ◽  
Direct Acting Antivirals ◽  
Resource Limited Settings ◽  
Rapid Diagnosis Test ◽  
Resource Limited ◽  
Specificity And Sensitivity ◽  
Hcv Screening ◽  
Direct Acting

Abstract Background Treatment of HCV with direct-acting antivirals has enabled the discussion of HCV eradication worldwide. Envisioning this aim requires implementation of mass screening in resource-limited areas, usually constrained by testing costs. Methods We validated a low-cost, rapid diagnosis test (RDT) for HCV in three different continents in 141 individuals. Results The HCV RDT showed 100% specificity and sensitivity across different samples regardless of genotype or viral load (in samples with such information, 90%). Conclusions The HCV test validated in this study can allow for HCV screening in areas of need when properly used.

scholarly journals Evaluation of Production Lots of a Rapid Point-of-Care Lateral Flow Serological Test Intended for Identification of IgM and IgG against the N-Terminal Part of the Spike Protein (S1) of SARS-CoV-2

Viruses ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 1043
Author(s):  
Tove Hoffman ◽  
Linda Kolstad ◽  
Bengt Rönnberg ◽  
Åke Lundkvist
Keyword(s):  
Predictive Value ◽  
Serological Test ◽  
Point Of Care ◽  
External Validation ◽  
High Sensitivity ◽  
Lateral Flow ◽  
Spike Protein ◽  
Individual Level ◽  
Lateral Flow Test ◽  
Production Errors

The potential of rapid point-of-care (POC) tests has been subject of doubt due to an eventual risk of production errors. The aim was therefore to evaluate the two separate production lots of a commercial POC lateral flow test, intended for the detection of IgM and IgG against the SARS-CoV-2 spike protein (S1). Control samples consisted of serum from individuals with confirmed SARS-CoV-2 infection and pre-COVID-19 negative sera gathered from a biobank. The presence of anti-S1 IgM/IgG in the sera was verified by an in-house Luminex-based serological assay (COVID-19 SIA). One hundred samples were verified as positive for anti-S1 IgG and 74 for anti-S1 IgM. Two hundred samples were verified as negative for anti-S1 IgM/IgG. For the two lots of the POC-test, the sensitivities were 93.2% and 87.8% for IgM and 93.0% and 100% for IgG. The specificities were 100% for IgM and 99.5% for IgG. The positive predictive value was 100% for IgM and 98.9% and 99.0% for IgG. The negative predictive value was 97.6% and 95.7% for IgM, and 96.6% and 100% for IgG. The evaluated POC-test is suitable to assess anti-SARS-CoV-2 S1 IgM and IgG, as a measure of previous virus exposure on an individual level. The external validation of separate lots of rapid POC-tests is encouraged to ensure high sensitivity before market introduction.

scholarly journals Rapid and Sensitive Detection of miRNA Based on AC Electrokinetic Capacitive Sensing for Point-of-Care Applications

Sensors ◽  
2021 ◽  
Vol 21 (12) ◽  
pp. 3985
Author(s):  
Nan Wan ◽  
Yu Jiang ◽  
Jiamei Huang ◽  
Rania Oueslati ◽  
Shigetoshi Eda ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Point Of Care ◽  
Low Cost ◽  
Clinical Diagnostics ◽  
Detection Methods ◽  
Capacitive Sensing ◽  
Application Potential ◽  
Mirna Detection ◽  
Mirna 25 ◽  
Low Sensitivity

A sensitive and efficient method for microRNAs (miRNAs) detection is strongly desired by clinicians and, in recent years, the search for such a method has drawn much attention. There has been significant interest in using miRNA as biomarkers for multiple diseases and conditions in clinical diagnostics. Presently, most miRNA detection methods suffer from drawbacks, e.g., low sensitivity, long assay time, expensive equipment, trained personnel, or unsuitability for point-of-care. New methodologies are needed to overcome these limitations to allow rapid, sensitive, low-cost, easy-to-use, and portable methods for miRNA detection at the point of care. In this work, to overcome these shortcomings, we integrated capacitive sensing and alternating current electrokinetic effects to detect specific miRNA-16b molecules, as a model, with the limit of detection reaching 1.0 femto molar (fM) levels. The specificity of the sensor was verified by testing miRNA-25, which has the same length as miRNA-16b. The sensor we developed demonstrated significant improvements in sensitivity, response time and cost over other miRNA detection methods, and has application potential at point-of-care.

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