iLluminate-miRNA: Paradigm for high-throughput, low-cost, and sensitive miRNA detection in serum samples at point-of-care

Prostate-specific antigen (PSA) is the established routine screening tool for the detection of early-stage prostate cancer. Given the laboratory-centric nature of the process, the development of a portable, ultra rapid high-throughput system for PSA screening is highly desirable. In this study, an advancedpoint-of-care system for PSA detection in human serum was developed based on a cellular biosensor where the cell membrane was modified by electroinserting a specific antibody against PSA. Thirty nine human serum samples were used for validation of this biosensory system for PSA detection. Samples were analyzed in parallel with a standard immunoradiometric assay (IRMA) and an established electrochemical immunoassay was used for comparison purposes. They were classified in three different PSA concentration ranges (0, <4 and ≥4 ng/mL). Cells membrane-engineered with 0.25 μg/mL anti-PSA antibody demonstrated a statistically lower response against the upper (≥4 ng/mL) PSA concentration range. In addition, the cell-based biosensor performed better than the immunosensor in terms of sensitivity and resolution against positive samples containing <4 ng/mL PSA. In spite of its preliminary, proof-of-concept stage of development, the cell-based biosensor could be used as aninitiative for the development of a fast, low-cost, and high-throughput POC screening system for PSA.

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

Sensors ◽

10.3390/s21123985 ◽

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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Field deployable platform for prognostic hepatic cancer screening.

Journal of Clinical Oncology ◽

10.1200/jco.2017.35.4_suppl.267 ◽

2017 ◽

Vol 35 (4_suppl) ◽

pp. 267-267

Author(s):

Jill E. Shea ◽

Jennifer H Granger ◽

Mark D Porter ◽

Courtney L. Scaife

Keyword(s):

High Risk ◽

Point Of Care ◽

Low Cost ◽

Serum Levels ◽

Blood Levels ◽

Vertical Flow ◽

Serum Samples ◽

Point Of Care System ◽

Risk Patients ◽

Care System

267 Background: Hepatocellular carcinoma (HCC) incidence rates are increasing in many parts of the world particularly in low- and middle-income countries (LMICs). Although treatable if identified in the early stages in many LMICs patients are identified later in disease. A low cost point of care system that could identify patients at a higher risk of having HCC could be used as a screening tool to identify patients at an earlier stage. We are currently developing a low cost point of care hand held platform that can simultaneously evaluate the blood levels of common HCC markers. We present our preliminary findings on the limit of detection (LOD) of our system. Methods: Our device combines the rapid but qualitative results of a vertical flow assay with the quantitative capabilities of surface enhanced Raman scattering spectroscopy. Our final product will be able to simultaneously measure the blood levels of the five most proven markers of HCC: alphafetoprotein (AFP), lens-culinaris agglutinin binding alphafetoprotein (AFP-L3), des-gamma-carboxy prothrombin (DCP), core antibodies to hepatitis B virus (HBV), and core antibodies to hepatitis C virus (HCV). In this pilot study we initially evaluated the LOD of our system by evaluating known concentrations of AFP in human serum samples. We then evaluated serum levels of AFP in HCC patient samples (n = 5) with our device and compared with values obtained using a standard ELISA. Results: The LOD of our system for the detection of AFP in spiked serum samples was 1 ng/mL. Our quantitative lateral flow assay measured AFP within 10% of the values of obtained by standard ELISA, with serum values ranging from 10-900 ng/mL. The quantitative vertical flow assay was able to provide results within 5 minutes at a cost of approximately $2/sample. Conclusions: Our inexpensive, portable and rapid test was able to measure serum levels of AFP at values comparable to standard clinical methods. Importantly, one blood marker would be insufficient to identify high risk patients, so further research will be required to expand the number of markers within the panel. The end goal will be to evaluate the ability of our point of care system to screen high risk population in LMICs, thereby identifying at risk patients earlier in disease progression.

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A Single-Bead-Based, Fully Integrated Microfluidic System for High-Throughput CD4+T Lymphocyte Enumeration

SLAS TECHNOLOGY Translating Life Sciences Innovation ◽

10.1177/2472630317737016 ◽

2017 ◽

Vol 23 (2) ◽

pp. 134-143 ◽

Cited By ~ 1

Author(s):

Xianbo Qiu ◽

Junhui Zhang ◽

Shisong Gong ◽

Dong Wang ◽

Shan Qiao ◽

...

Keyword(s):

T Lymphocyte ◽

High Throughput ◽

Point Of Care ◽

Low Cost ◽

Ccd Camera ◽

Microfluidic System ◽

Magnetic Control ◽

Magnetic Actuation ◽

Fully Integrated ◽

Single Bead

A single-bead-based, fully integrated microfluidic system has been developed for high-throughput CD4+T lymphocyte enumeration at point-of-care testing. Instead of directly counting CD4+T lymphocytes, CD4+T lymphocyte enumeration is achieved by quantitatively detecting CD4 antigen from the lysed blood sample with a functionalized polycarbonate single bead based on chemiluminescence. To implement the sandwiched chemiluminescence immunoassay with reduced nonspecific binding, a streamlined microfluidic chip with multiple reaction chambers is developed to allow each reaction step to be completed in an independent chamber where reagent is pre-stored. With simple magnetic control, the single bead with an embedded ferrous core can be consecutively transported between each of two adjacent chambers for different reactions. Meanwhile, enhanced mixing can be achieved by moving the single bead back and forth inside one chamber with magnetic actuation. High-throughput detection can be performed when a linear actuation stage is adopted to introduce synchronous magnetic control to multiple single beads in parallel microfluidic chips. A sensitive charge-coupled device (CCD) camera is adopted for high-throughput chemiluminescence detection from multiple single beads. Experimental results show that with the fully integrated microfluidic system, easy-to-operate, accurate, low-cost, immediate, and high-throughput CD4+T lymphocyte enumeration can be successfully achieved at resource-poor settings.

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Deep Learning-Enabled Point-of-Care Sensing Using Multiplexed Paper-Based Sensors

10.1101/667436 ◽

2019 ◽

Cited By ~ 1

Author(s):

Zachary Ballard ◽

Hyou-Arm Joung ◽

Artem Goncharov ◽

Jesse Liang ◽

Karina Nugroho ◽

...

Keyword(s):

Machine Learning ◽

Deep Learning ◽

Point Of Care ◽

Low Cost ◽

High Sensitivity ◽

Cost Effective ◽

Serum Samples ◽

Cvd Risk ◽

Medical Test ◽

Sensing Membrane

ABSTRACTWe present a deep learning-based framework to design and quantify point-of-care sensors. As its proof-of-concept and use-case, we demonstrated a low-cost and rapid paper-based vertical flow assay (VFA) for high sensitivity C-Reactive Protein (hsCRP) testing, a common medical test used for quantifying the degree of inflammation in patients at risk of cardio-vascular disease (CVD). A machine learning-based sensor design framework was developed for two key tasks: (1) to determine an optimal configuration of immunoreaction spots and conditions, spatially-multiplexed on a paper-based sensing membrane, and (2) to accurately infer the target analyte concentration based on the signals of the optimal VFA configuration. Using a custom-designed mobile-phone based VFA reader, a clinical study was performed with 85 human serum samples to characterize the quantification accuracy around the clinically defined cutoffs for CVD risk stratification. Results from blindly-tested VFAs indicate a competitive coefficient of variation of 11.2% with a linearity of R2 = 0.95; in addition to the success in the high-sensitivity CRP range (i.e., 0-10 mg/L), our results further demonstrate a mitigation of the hook-effect at higher CRP concentrations due to the incorporation of antigen capture spots within the multiplexed sensing membrane of the VFA. This paper-based computational VFA that is powered by deep learning could expand access to CVD health screening, and the presented machine learning-enabled sensing framework can be broadly used to design cost-effective and mobile sensors for various point-of-care diagnostics applications.

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Advances in Fully Automated Nucleic Acid Extraction System Based on Magnetic Nanobeads

Nanoscience and Nanotechnology Letters ◽

10.1166/nnl.2019.3051 ◽

2019 ◽

Vol 11 (12) ◽

pp. 1633-1643

Author(s):

Zhu Chen ◽

Changhu Xiao ◽

Manling Tang ◽

Yuyue Xu ◽

Ziyu He ◽

...

Keyword(s):

Nucleic Acid ◽

High Throughput ◽

Point Of Care ◽

Low Cost ◽

Future Trend ◽

Acid Extraction ◽

Nucleic Acid Extraction ◽

Advantages And Disadvantages ◽

Good Repeatability ◽

Magnetic Nanobeads

Using magnetic nanobeads (MNBs) to extract nucleic acids is an efficient, inexpensive, easy automation, high throughput and good repeatability, instead of the traditional nucleic acid extraction (NAE) methods. Advances in fully automated MNBs-based nucleic acid extraction systems (MNAES) can push the frontiers of point-of-care testing (POCT) devices towards low-cost, automation, and enhanced accuracy molecular-level diagnostics. So, this paper introduces the pipettingbased MNAES with position of magnetic separation kit or tip, and based on magnetic bar MNAES with blending manner is shock or rotation. Further, advantages and disadvantages of various MNAES are compared. We envisage that the future trend in molecular diagnosis and monitoring will be facilitation, intelligent, miniaturization, and high throughput MNAES with sample-in-answer-out capability.

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Detection and quantification of Mycobacterium tuberculosis antigen CFP10 in serum and urine for the rapid diagnosis of active tuberculosis disease

Scientific Reports ◽

10.1038/s41598-021-98471-1 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Marva Seifert ◽

Eva Vargas ◽

Victor Ruiz-Valdepeñas Montiel ◽

Joseph Wang ◽

Timothy C. Rodwell ◽

...

Keyword(s):

Point Of Care ◽

Low Cost ◽

Specific Antigen ◽

Active Tuberculosis ◽

Serum Samples ◽

Tuberculosis Patients ◽

Tuberculosis Antigen ◽

Highly Sensitive ◽

Tuberculosis Disease ◽

Serum And Urine

AbstractOutside of the ongoing COVID-19 pandemic, tuberculosis is the leading cause of infectious disease mortality globally. Currently, there is no commercially available point-of-care diagnostic that is rapid, inexpensive, and highly sensitive for the diagnosis of active tuberculosis disease. Here we describe the development and optimization of a novel, highly sensitive prototype bioelectronic tuberculosis antigen (BETA) assay to detect tuberculosis-specific antigen, CFP10, in small-volume serum and urine samples. In this proof-of-concept study we evaluated the performance of the BETA assay using clinical specimens collected from presumptive tuberculosis patients from three independent cohorts. Circulating CFP10 antigen was detected in ALL serum (n = 19) and urine (n = 3) samples from bacteriologically confirmed tuberculosis patients who were untreated or had less than one week of treatment at time of serum collection, successfully identifying all culture positive tuberculosis patients. No CFP10 antigen was detected in serum (n = 7) or urine (n = 6) samples from individuals who were determined to be negative for tuberculosis disease. Additionally, antigen quantification using the BETA assay of paired serum samples collected from tuberculosis patients (n = 8) both before and after treatment initiation, indicate consistently declining within-person levels of CFP10 antigen during treatment. This novel, low-cost assay demonstrates potential as a rapid, non-sputum-based, point-of-care tool for the diagnosis of tuberculosis disease.

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Lateral Flow Aptasensor for Simultaneous Detection of Platelet-Derived Growth Factor-BB (PDGF-BB) and Thrombin

Molecules ◽

10.3390/molecules24040756 ◽

2019 ◽

Vol 24 (4) ◽

pp. 756 ◽

Cited By ~ 2

Author(s):

Guodong Liu ◽

Anant Gurung ◽

Wanwei Qiu

Keyword(s):

Growth Factor ◽

Point Of Care ◽

Low Cost ◽

Simultaneous Detection ◽

Lateral Flow ◽

Platelet Derived Growth Factor ◽

Great Promise ◽

Serum Samples ◽

Lateral Flow Strip ◽

Assay Time

Here we report a lateral flow aptasensor (LFA) for the simultaneous detection of platelet-derived growth factor-BB (PDGF-BB) and thrombin. Two pairs of aptamers, which are specific against PDGF-BB and thrombin, respectively, were used to prepare the LFA. Thiolated aptamers were immobilized on a gold nanoparticle (AuNP) surface and biotinylated aptamers were immobilized on the test zones of an LFA nitrocellulose membrane. The assay involved the capture of PDGF-BB and thrombin simultaneously in sandwich-type formats between the capture aptamers on the test zones of LFA and AuNP-labeled detection aptamers. AuNPs were thus captured on the test zones of the LFA and gave red bands to enable the visual detection of target proteins. Quantitative results were obtained by reading the test band intensities with a portable strip reader. By combining the highly specific molecular recognition properties of aptamers with the unique properties of lateral flow assay (low-cost, short assay time and a user-friendly format), the optimized aptasensor was capable of simultaneously detecting 1.0 nM of PDGF-BB and 1.5 nM of thrombin in association with a 10-min assay time. The biosensor was also successfully applied to detect PDGF-BB and thrombin in spiked human serum samples. The LFA shows great promise for the development of aptamer-based lateral flow strip biosensors for point-of-care or for the in-field detection of disease-related protein biomarkers.

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OC 8435 MULTI-BIOMARKER TEST STRIP FOR POINT-OF-CARE SCREENING FOR ACTIVE TUBERCULOSIS: A FIVE-COUNTRY MULTI-CENTRE TEST EVALUATION

BMJ Global Health ◽

10.1136/bmjgh-2019-edc.14 ◽

2019 ◽

Vol 4 (Suppl 3) ◽

pp. A6.2-A6

Author(s):

Paul Corstjens ◽

Anouk Van Hooij ◽

Elisa Tjon Kon Fat ◽

Shannon Herdigein ◽

Anna Ritah Namuganga ◽

...

Keyword(s):

Respiratory Diseases ◽

Serum Proteins ◽

Point Of Care ◽

Low Cost ◽

Test Strip ◽

Screening Tests ◽

Rapid Screening ◽

Serum Samples ◽

Multiple Test ◽

Biomarker Selection

BackgroundInexpensive rapid screening tests that can be used at the point-of-care (POC) are vital to combat tuberculosis. Particularly, less invasive non-sputum-based biomarker tests for all TB forms can help controlling transmission. Availability of such tests would significantly accelerate and streamline diagnostic approaches, improve cost-efficiency and decrease unnecessary costly GeneXpert referrals.MethodsMulti-biomarker test (MBT) devices measuring levels of selections of up to six serum proteins simultaneously on a single lateral flow (LF) strip were produced. The strip contains individual capture lines for a biomarker selection allowing discrimination of TB-patients from other respiratory diseases (ORD). Only biomarkers successfully evaluated with singleplex strips (single biomarker tests) were applied to the MBT device. Quantitative signals are recorded with a low-cost handheld reader compatible with the applied luminescent up-converting particle (UCP) label. Biomarker selection and algorithms used to distinguish potential-TB and ORD are flexible.ResultsResults obtained with MBT strips containing multiple test lines correlate well with singleplex LF strips. Using LF tests for 5 selected biomarkers a sensitivity of 94% and specificity of 96% could be achieved with a confirmed South African selection of 20 TB and 31 non-TB samples. Patients were designated TB positive when scoring a value above the cut-off threshold for at least 3 out of 5 biomarkers. Serum samples of potential TB patients collected at five medical research institutes (Ethiopia, Namibia, South Africa, The Gambia, Uganda) were tested locally with MBT strips comprised of CRP, SAA, IP-10, Ferritin, ApoA-I and IL-6 and results analysed to obtain an overall pan-Africa applicable signature.ConclusionEvaluated POC applicable UCP-LF devices detecting serum biomarker signatures can help to distinguish active TB from other respiratory diseases and as such can prioritise highest-risk patients for further care. Ongoing prospective studies evaluate the MBT strip with fingerstick blood and do not require a laboratory or trained phlebotomist anymore.

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Expanding the Toolbox for Label-Free Enzyme Assays: A Dinuclear Platinum(II) Complex/DNA Ensemble with Switchable Near-IR Emission

Molecules ◽

10.3390/molecules24234390 ◽

2019 ◽

Vol 24 (23) ◽

pp. 4390 ◽

Cited By ~ 1

Author(s):

Moustafa T. Gabr ◽

F. Christopher Pigge

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Low Cost ◽

Dnase I ◽

Dna Assembly ◽

Label Free ◽

Serum Samples ◽

Near Ir ◽

Dinuclear Platinum ◽

Label Free Detection

Switchable luminescent bioprobes whose emission can be turned on as a function of specific enzymatic activity are emerging as important tools in chemical biology. We report a promising platform for the development of label-free and continuous enzymatic assays in high-throughput mode based on the reversible solvent-induced self-assembly of a neutral dinuclear Pt(II) complex. To demonstrate the utility of this strategy, the switchable luminescence of a dinuclear Pt(II) complex was utilized in developing an experimentally simple, fast (10 min), low cost, and label-free turn-on luminescence assay for the endonuclease enzyme DNAse I. The complex displays a near-IR (NIR) aggregation-induced emission at 785 nm in aqueous solution that is completely quenched upon binding to G-quadruplex DNA from the human c-myc oncogene. Luminescence is restored upon DNA degradation elicited by exposure to DNAse I. Correlation between near-IR luminescence intensity and DNAse I concentration in human serum samples allows for fast and label-free detection of DNAse I down to 0.002 U/mL. The Pt(II) complex/DNA assembly is also effective for identification of DNAse I inhibitors, and assays can be performed in multiwell plates compatible with high-throughput screening. The combination of sensitivity, speed, convenience, and cost render this method superior to all other reported luminescence-based DNAse I assays. The versatile response of the Pt(II) complex to DNA structures promises broad potential applications in developing real-time and label-free assays for other nucleases as well as enzymes that regulate DNA topology.

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