Companion and Point-of-Care Sensor System for Rapid Multiplexed Detection of a Panel of Infectious Disease Markers

2017 ◽  
pp. 247263031769677
Author(s):  
Anjan Panneer Selvam ◽  
Shalini Prasad
Keyword(s):  
Whole Blood ◽  
Dynamic Range ◽  
Electrochemical Impedance ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Lipoteichoic Acid ◽  
Impedance Change ◽  
Label Free ◽  
Nylon Membrane ◽  
Multiplexed Detection

A nanochannel-based electrochemical biosensor has been demonstrated for rapid and multiplexed detection of a panel of three biomarkers associated with rapid detection of sepsis. The label-free biosensor detected procalcitonin (PCT), lipoteichoic acid (LTA), and lipopolysaccharide (LPS) from human whole blood. The biosensor comprises a nanoporous nylon membrane integrated onto a microelectrode sensor platform for nanoconfinement effects. Charge perturbations due to biomarker binding are recorded as impedance changes using electrochemical impedance spectroscopy. The measured impedance change is used to quantitatively determine the concentration of the three biomarkers using antibody receptors from the tested sample. We were successful in detecting and quantifying the three biomarkers from whole blood. The limit of detection was 0.1 ng/mL for PCT and 1 µg/mL for LPS and LTA. The sensor was able to demonstrate a dynamic range of detection from 01.1 ng/mL to 10 µg/mL for PCT and from 1 µg/mL to 1000 µg/mL for LPS and LTA biomarkers. This novel technology has promising preliminary results toward the design of sensors for rapid and sensitive detection of the three panel biomarkers in whole blood toward diagnosis and classification of sepsis.

scholarly journals Companion and Point-of-Care Sensor System for Rapid Multiplexed Detection of a Panel of Infectious Disease Markers

2017 ◽  
Vol 22 (3) ◽  
pp. 338-347 ◽  
Author(s):  
Anjan Panneer Selvam ◽  
Shalini Prasad
Keyword(s):  
Whole Blood ◽  
Dynamic Range ◽  
Electrochemical Impedance ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Lipoteichoic Acid ◽  
Impedance Change ◽  
Label Free ◽  
Nylon Membrane ◽  
Multiplexed Detection

A nanochannel-based electrochemical biosensor has been demonstrated for rapid and multiplexed detection of a panel of three biomarkers associated with rapid detection of sepsis. The label-free biosensor detected procalcitonin (PCT), lipoteichoic acid (LTA), and lipopolysaccharide (LPS) from human whole blood. The biosensor comprises a nanoporous nylon membrane integrated onto a microelectrode sensor platform for nanoconfinement effects. Charge perturbations due to biomarker binding are recorded as impedance changes using electrochemical impedance spectroscopy. The measured impedance change is used to quantitatively determine the concentration of the three biomarkers using antibody receptors from the tested sample. We were successful in detecting and quantifying the three biomarkers from whole blood. The limit of detection was 0.1 ng/mL for PCT and 1 µg/mL for LPS and LTA. The sensor was able to demonstrate a dynamic range of detection from 01.1 ng/mL to 10 µg/mL for PCT and from 1 µg/mL to 1000 µg/mL for LPS and LTA biomarkers. This novel technology has promising preliminary results toward the design of sensors for rapid and sensitive detection of the three panel biomarkers in whole blood toward diagnosis and classification of sepsis.

scholarly journals Label Free, Lateral Flow Prostaglandin E2 Electrochemical Immunosensor for Urinary Tract Infection Diagnosis

Chemosensors ◽  
2021 ◽  
Vol 9 (9) ◽  
pp. 271
Author(s):  
Antra Ganguly ◽  
Tahmineh Ebrahimzadeh ◽  
Philippe E. Zimmern ◽  
Nicole J. De Nisco ◽  
Shalini Prasad
Keyword(s):  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Tract Infection ◽  
Prostaglandin E2 ◽  
Dynamic Range ◽  
Electrochemical Impedance ◽  
Point Of Care ◽  
Treatment Success ◽  
Lateral Flow ◽  
Label Free

A label-free, rapid, and easy-to-use lateral flow electrochemical biosensor was developed for urinary tract infection (UTI) diagnosis in resource challenged areas. The sensor operates in non-faradaic mode and utilizes Electrochemical Impedance Spectroscopy for quantification of Prostaglandin E2, a diagnostic and prognostic urinary biomarker for UTI and recurrent UTI. To achieve high sensitivity in low microliter volumes of neat, unprocessed urine, nanoconfinement of assay biomolecules was achieved by developing a three-electrode planar gold microelectrode system on top of a lateral flow nanoporous membrane. The sensor is capable of giving readouts within 5 min and has a wide dynamic range of 100–4000 pg/mL for urinary PGE2. The sensor is capable of discriminating between low and high levels of PGE2 and hence is capable of threshold classification of urine samples as UTI positive and UTI negative. The sensor through its immunological response (directly related to host immune response) is superior to the commercially available point-of-care UTI dipsticks which are qualitative, have poor specificity for UTI, and have high false-positive rates. The developed sensor shows promise for rapid, easy and cost-effective UTI diagnosis for both clinical and home-based settings. More accurate point-of-care UTI diagnosis will improve patient outcomes and allow for timely and appropriate prescription of antibiotics which can subsequently increase treatment success rates and reduce costs.

scholarly journals Sensitive Detection of Francisella tularensis Directly from Whole Blood by Use of the GeneXpert System

2016 ◽  
Vol 55 (1) ◽  
pp. 291-301 ◽  
Author(s):  
Padmapriya P. Banada ◽  
Srinidhi Deshpande ◽  
Soumitesh Chakravorty ◽  
Riccardo Russo ◽  
James Occi ◽  
...  
Keyword(s):  
Blood Culture ◽  
Francisella Tularensis ◽  
Whole Blood ◽  
Dynamic Range ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Bloodstream Infections ◽  
Content Type ◽  
Testing Procedures ◽  
Point Of Care Detection

ABSTRACTFrancisella tularensisis a potential bioterrorism agent that is highly infectious at very low doses. Diagnosis of tularemia by blood culture and nucleic acid-based diagnostic tests is insufficiently sensitive. Here, we demonstrate a highly sensitiveF. tularensisassay that incorporates sample processing and detection into a single cartridge suitable for point-of-care detection. The assay limit of detection (LOD) and dynamic range were determined in a filter-based cartridge run on the GeneXpert system.F. tularensisDNA in buffer or CFU ofF. tularensiswas spiked into human or macaque blood. To simulate detection in human disease, the assay was tested on blood drawn from macaques infected withF. tularensisSchu S4 at daily intervals. Assay detection was compared to that with a conventional quantitative PCR (qPCR) assay and blood culture. The assay LOD was 0.1 genome equivalents (GE) per reaction and 10 CFU/mlF. tularensisin both human and macaque blood. In infected macaques, the assay detectedF. tularensison days 1 to 4 postinfection in 21%, 17%, 60%, and 83% of macaques, respectively, compared to conventional qPCR positivity rates of 0%, 0%, 30%, and 100% and CFU detection of blood culture at 0%, 0%, 0%, and 10% positive, respectively. Assay specificity was 100%. The new cartridge-based assay can rapidly detectF. tularensisin bloodstream infections directly in whole blood at the early stages of infection with a sensitivity that is superior to that of other methods. The simplicity of the automated testing procedures may make this test suitable for rapid point-of-care detection.

scholarly journals Label free, electric field mediated ultrasensitive electrochemical point-of-care device for CEA detection

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
B. Chakraborty ◽  
A. Das ◽  
N. Mandal ◽  
N. Samanta ◽  
N. Das ◽  
...  
Keyword(s):  
Electric Field ◽  
Electrochemical Impedance ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Low Cost ◽  
Zno Nanorods ◽  
Hybrid Nanostructures ◽  
Electrochemical Sensing ◽  
Label Free ◽  
Electron Transfer Rate

AbstractDeveloping point-of-care (PoC) diagnostic platforms for carcinoembryonic antigen detection is essential. However, thefew implementations of transferring the signal amplification strategies in electrochemical sensing on paper-based platforms are not satisfactory in terms of detection limit (LOD). In the quest for pushing down LOD, majority of the research has been targeted towards development of improved nanostructured substrates for entrapping more analyte molecules and augmenting the electron transfer rate to the working electrode. But, such approaches have reached saturation. This paper focuses on enhancing the mass transport of the analyte towards the sensor surface through the application of an electric field, in graphene-ZnO nanorods heterostructure. These hybrid nanostructures have been deposited on flexible polyethylene terephthalate substrates with screen printed electrodes for PoC application. The ZnO nanorods have been functionalized with aptamers and the working sensor has been integrated with smartphone interfaced indigenously developed low cost potentiostat. The performance of the system, requiring only 50 µl analyte has been evaluated using electrochemical impedance spectroscopy and validated against commercially available ELISA kit. Limit of detection of 1 fg/ml in human serum with 6.5% coefficient of variation has been demonstrated, which is more than three orders of magnitude lower than the existing attempts on PoC device.

scholarly journals PERFORMANCE CHARACTERISTICS OF A FRET-BASED IMMUNOASSAY FOR QUANTITATION OF INFLIXIMAB ON A POINT-OF-CARE INSTRUMENT SYSTEM

2021 ◽  
Vol 27 (Supplement_1) ◽  
pp. S57-S57
Author(s):  
Edgar Ong ◽  
Ruo Huang ◽  
Richard Kirkland ◽  
Michael Hale ◽  
Larry Mimms
Keyword(s):  
Whole Blood ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Quantitative Detection ◽  
Therapeutic Drug ◽  
Analytical Sensitivity ◽  
Sample Type ◽  
Hook Effect ◽  
Limit Of Quantitation ◽  
Assay Precision

Abstract Introduction A fast (<5 min), time-resolved fluorescence resonance energy transfer (FRET)-based immunoassay was developed for the quantitative detection of infliximab (IFX) and biosimilars for use in therapeutic drug monitoring using only 20 µL of fingerstick whole blood or serum at the point-of-care. The Procise IFX assay and ProciseDx analyzer are CE-marked. Studies were performed to characterize analytical performance of the Procise IFX assay on the ProciseDx analyzer. Methods Analytical testing was performed by spiking known amounts of IFX into negative serum and whole blood specimens. Analytical sensitivity was determined using limiting concentrations of IFX. Linearity was determined by testing IFX across the assay range. Hook effect was assessed at IFX concentrations beyond levels expected to be found within a patient. Testing of assay precision, cross-reactivity and potential interfering substances, and biosimilars was performed. The Procise IFX assay was also compared head-to-head with another CE-marked assay: LISA-TRACKER infliximab ELISA test (Theradiag, France). The accuracy of the Procise IFX assay is established through calibrators and controls traceable to the WHO 1st International Standard for Infliximab (NIBSC code: 16/170). Results The Procise IFX assay shows a Limit of Blank, Limit of Detection, and Lower Limit of Quantitation (LLoQ) of 0.1, 0.2, and 1.1 µg/mL in serum and 0.6, 1.1, and 1.7 µg/mL in whole blood, respectively. The linear assay range was determined to be 1.7 to 77.2 µg/mL in serum and whole blood. No hook effect was observed at an IFX concentration of 200 µg/mL as the value reported as “>ULoQ”. Assay precision testing across 20 days with multiple runs and reagent lots showed an intra-assay coefficient of variation (CV) of 2.7%, an inter-assay CV of <2%, and a total CV of 3.4%. The presence of potentially interfering/cross-reacting substances showed minimal impact on assay specificity with %bias within ±8% of control. Testing of biosimilars (infliximab-dyyb and infliximab-abda) showed good recovery. A good correlation to the Theradiag infliximab ELISA was obtained for both serum (slope=1.01; r=0.99) and whole blood (slope=1.01; r=0.98) samples (Figure 1). Conclusion Results indicate that the Procise IFX assay is sensitive, specific, and precise yielding results within 5 minutes from both whole blood and serum without the operator needing to specify sample type. Additionally, it shows very good correlation to a comparator assay that takes several hours and sample manipulation to yield results. This makes the Procise IFX assay ideal for obtaining fast and accurate IFX quantitation, thus allowing for immediate drug level dosing decisions to be made by the physician during patient treatment.

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 Label-Free Electrochemical Sensor for Rapid Bacterial Pathogen Detection Using Vancomycin-Modified Highly Branched Polymers

Sensors ◽  
2021 ◽  
Vol 21 (5) ◽  
pp. 1872
Author(s):  
Holger Schulze ◽  
Harry Wilson ◽  
Ines Cara ◽  
Steven Carter ◽  
Edward N. Dyson ◽  
...  
Keyword(s):  
Pathogen Detection ◽  
Electrochemical Impedance ◽  
Point Of Care ◽  
Branched Polymers ◽  
Label Free ◽  
Conformation Change ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
Staphylococcus Carnosus ◽  
Label Free Detection

Rapid point of care tests for bacterial infection diagnosis are of great importance to reduce the misuse of antibiotics and burden of antimicrobial resistance. Here, we have successfully combined a new class of non-biological binder molecules with electrochemical impedance spectroscopy (EIS)-based sensor detection for direct, label-free detection of Gram-positive bacteria making use of the specific coil-to-globule conformation change of the vancomycin-modified highly branched polymers immobilized on the surface of gold screen-printed electrodes upon binding to Gram-positive bacteria. Staphylococcus carnosus was detected after just 20 min incubation of the sample solution with the polymer-functionalized electrodes. The polymer conformation change was quantified with two simple 1 min EIS tests before and after incubation with the sample. Tests revealed a concentration dependent signal change within an OD600 range of Staphylococcus carnosus from 0.002 to 0.1 and a clear discrimination between Gram-positive Staphylococcus carnosus and Gram-negative Escherichia coli bacteria. This exhibits a clear advancement in terms of simplified test complexity compared to existing bacteria detection tests. In addition, the polymer-functionalized electrodes showed good storage and operational stability.

scholarly journals Highly sensitive dual mode electrochemical platform for microRNA detection

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Pawan Jolly ◽  
Marina R. Batistuti ◽  
Anna Miodek ◽  
Pavel Zhurauski ◽  
Marcelo Mulato ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Dynamic Range ◽  
Electrochemical Impedance ◽  
Limit Of Detection ◽  
Dual Mode ◽  
Electrode Surfaces ◽  
Microrna Detection ◽  
Highly Sensitive ◽  
Detection Mode ◽  
Amplification Strategy

Abstract MicroRNAs (miRNAs) play crucial regulatory roles in various human diseases including cancer, making them promising biomarkers. However, given the low levels of miRNAs present in blood, their use as cancer biomarkers requires the development of simple and effective analytical methods. Herein, we report the development of a highly sensitive dual mode electrochemical platform for the detection of microRNAs. The platform was developed using peptide nucleic acids as probes on gold electrode surfaces to capture target miRNAs. A simple amplification strategy using gold nanoparticles has been employed exploiting the inherent charges of the nucleic acids. Electrochemical impedance spectroscopy was used to monitor the changes in capacitance upon any binding event, without the need for any redox markers. By using thiolated ferrocene, a complementary detection mode on the same sensor was developed where the increasing peaks of ferrocene were recorded using square wave voltammetry with increasing miRNA concentration. This dual-mode approach allows detection of miRNA with a limit of detection of 0.37 fM and a wide dynamic range from 1 fM to 100 nM along with clear distinction from mismatched target miRNA sequences. The electrochemical platform developed can be easily expanded to other miRNA/DNA detection along with the development of microarray platforms.

scholarly journals Multitarget, quantitative nanoplasmonic electrical field-enhanced resonating device (NE2RD) for diagnostics

2015 ◽  
Vol 112 (32) ◽  
pp. E4354-E4363 ◽  
Author(s):  
Fatih Inci ◽  
Chiara Filippini ◽  
Murat Baday ◽  
Mehmet Ozgun Ozen ◽  
Semih Calamak ◽  
...  
Keyword(s):  
Dynamic Range ◽  
Point Of Care ◽  
Medical Diagnostics ◽  
Clinical Samples ◽  
Label Free ◽  
Protein Biomarkers ◽  
Electrical Field ◽  
Color Changes ◽  
Sensing Platform ◽  
Broad Dynamic Range

Recent advances in biosensing technologies present great potential for medical diagnostics, thus improving clinical decisions. However, creating a label-free general sensing platform capable of detecting multiple biotargets in various clinical specimens over a wide dynamic range, without lengthy sample-processing steps, remains a considerable challenge. In practice, these barriers prevent broad applications in clinics and at patients’ homes. Here, we demonstrate the nanoplasmonic electrical field-enhanced resonating device (NE2RD), which addresses all these impediments on a single platform. The NE2RD employs an immunodetection assay to capture biotargets, and precisely measures spectral color changes by their wavelength and extinction intensity shifts in nanoparticles without prior sample labeling or preprocessing. We present through multiple examples, a label-free, quantitative, portable, multitarget platform by rapidly detecting various protein biomarkers, drugs, protein allergens, bacteria, eukaryotic cells, and distinct viruses. The linear dynamic range of NE2RD is five orders of magnitude broader than ELISA, with a sensitivity down to 400 fg/mL This range and sensitivity are achieved by self-assembling gold nanoparticles to generate hot spots on a 3D-oriented substrate for ultrasensitive measurements. We demonstrate that this precise platform handles multiple clinical samples such as whole blood, serum, and saliva without sample preprocessing under diverse conditions of temperature, pH, and ionic strength. The NE2RD’s broad dynamic range, detection limit, and portability integrated with a disposable fluidic chip have broad applications, potentially enabling the transition toward precision medicine at the point-of-care or primary care settings and at patients’ homes.

scholarly journals Whole Blood Immunoassay Based on Centrifugal Bead Sedimentation

2011 ◽  
Vol 57 (5) ◽  
pp. 753-761 ◽  
Author(s):  
Ulrich Y Schaff ◽  
Greg J Sommer
Keyword(s):  
Whole Blood ◽  
Single Channel ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Low Cost ◽  
Microfluidic System ◽  
Blood Samples ◽  
Reactive Protein ◽  
Immunoassay Technique ◽  
Microfluidic Immunoassay

BACKGROUND Centrifugal “lab on a disk” microfluidics is a promising avenue for developing portable, low-cost, automated immunoassays. However, the necessity of incorporating multiple wash steps results in complicated designs that increase the time and sample/reagent volumes needed to run assays and raises the probability of errors. We present proof of principle for a disk-based microfluidic immunoassay technique that processes blood samples without conventional wash steps. METHODS Microfluidic disks were fabricated from layers of patterned, double-sided tape and polymer sheets. Sample was mixed on-disk with assay capture beads and labeling antibodies. Following incubation, the assay beads were physically separated from the blood cells, plasma, and unbound label by centrifugation through a density medium. A signal-laden pellet formed at the periphery of the disk was analyzed to quantify concentration of the target analyte. RESULTS To demonstrate this technique, the inflammation biomarkers C-reactive protein and interleukin-6 were measured from spiked mouse plasma and human whole blood samples. On-disk processing (mixing, labeling, and separation) facilitated direct assays on 1-μL samples with a 15-min sample-to-answer time, <100 pmol/L limit of detection, and 10% CV. We also used a unique single-channel multiplexing technique based on the sedimentation rate of different size or density bead populations. CONCLUSIONS This portable microfluidic system is a promising method for rapid, inexpensive, and automated detection of multiple analytes directly from a drop of blood in a point-of-care setting.

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