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1.
Heliyon ; 10(18): e37722, 2024 Sep 30.
Article in English | MEDLINE | ID: mdl-39328528

ABSTRACT

Most urine test strips are intended to enable the general population to rapidly and easily diagnose potential renal disorders. It is semi-quantitative in nature, and although the procedure is straightforward, certain factors will affect the judgmental outcomes. This study describes rapid and accurate quantification of twelve urine test strip parameters: leukocytes, nitrite, urobilinogen, protein, pH, occult blood, specific gravity, ketone, bilirubin, glucose, microalbumin, and creatinine using a micro-electromechanical system (MEMS)-based spectrophotometer, known as a spectrochip. For each parameter, absorption spectra were measured three times independently at eight different concentration levels of diluted standard solutions, and the average spectral intensities were calculated to establish the calibration curve under the characteristic wavelength ( λ c ). Then, regression analysis on the calibration curve was performed with GraphPad Prism software, which revealed that the coefficient of determination ( R 2 ) of the modeled calibration curves was greater than 0.95. This result illustrates that the measurements exceed standard levels, confirming the importance of a spectrochip for routine multi-parameter urine analysis. Thus, it is possible to obtain the spectral signal strength for each parameter at its characteristic wavelength in order to compare directly with the calibration curves in the future, even in situations when sample concentration is unknown. Additionally, the use of large testing machines can be reduced in terms of cost, time, and space by adopting a micro urine testing platform based on spectrochip, which also improves operational convenience and effectively enables point-of-care (POC) testing in urinalysis.

2.
Iran J Biotechnol ; 22(2): e3722, 2024 Apr.
Article in English | MEDLINE | ID: mdl-39220332

ABSTRACT

Background: The early detection of infectious microorganisms is crucial for preventing and controlling the transmission of diseases. This article provides a comprehensive review of biosensors based on various diagnostic methods for measuring airborne pathogens. Objective: This article aims to explore recent advancements in the field of biosensors tailored for the detection and monitoring of airborne microorganisms, offering insights into emerging technologies and their potential applications in environmental surveillance and public health management. Materials and Methods: The study summarizes the research conducted on novel methods of detecting airborne microorganisms using different biological sensors, as well as the application of signal amplification technologies such as polymerase chain reaction (PCR), immunoassay reactions, molecular imprinted polymers (MIP) technique, lectin and cascade reactions, and nanomaterials. Results: Antibody and PCR detection methods are effective for specific microbial strains, but they have limitations including limited stability, high cost, and the need for skilled operators with basic knowledge of the target structure. Biosensors based on MIP and lectin offer a low-cost, stable, sensitive, and selective alternative to antibodies and PCR. However, challenges remain, such as the detection of small gas molecules by MIP and the lower sensitivity of lectins compared to antibodies. Additionally, achieving high sensitivity in complex environments poses difficulties for both methods. Conclusion: The development of sensitive, reliable, accessible, portable, and inexpensive biosensors holds great potential for clinical and environmental applications, including disease diagnosis, treatment monitoring, and point-of-care testing, offering a promising future in this field. This review presents an overview of biosensor detection principles, covering component identification, energy conversion principles, and signal amplification. Additionally, it summarizes the research and applications of biosensors in the detection of airborne microorganisms. The latest advancements and future trends in biosensor detection of airborne microorganisms are also analyzed.

3.
Expert Rev Mol Diagn ; 24(9): 829-839, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39135402

ABSTRACT

INTRODUCTION: Sepsis is a heterogeneous syndrome often misdiagnosed. Point-of-care (POC) diagnostic tests are commonly used to guide decision and include host biomarkers and molecular diagnostics. AREAS COVERED: The diagnostic and prognostic accuracy of established and emerging biomarkers for sepsis, including procalcitonin (PCT) soluble urokinase plasminogen activator receptor (suPAR), presepsin, TRAIL/IP-10/CRP, MxA, and MxA-CRP, are analyzed in this review. The clinical utility of the two prevalent molecular techniques for pathogens identification using polymerase chain reaction (PCR) assays is also presented: FILMARRAY and QIAstat-Dx RP. EXPERT OPINION: The rising benefits of the combined use of POC biomarkers with molecular diagnostics in daily clinical routine appear to outperform conventional practices in terms of reduced turnaround time, timely diagnosis, and prompt administration of the appropriate treatment. Yet, this must be further demonstrated in future investigations. However, the cost-effectiveness of POC tests and the high rate of false positive and negative results, indicate the need for a comprehensive clinical evaluation.


Subject(s)
Biomarkers , Point-of-Care Testing , Sepsis , Humans , Biomarkers/blood , Sepsis/diagnosis , Sepsis/blood , Molecular Diagnostic Techniques/methods , Receptors, Urokinase Plasminogen Activator/blood , Procalcitonin/blood , Point-of-Care Systems
4.
Adv Mater ; 36(39): e2311931, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39086075

ABSTRACT

Polymerase chain reaction (PCR) is the gold standard for nucleic acid amplification and quantification in diverse fields such as life sciences, global health, medicine, agricultural science, forensic science, and environmental science for global sustainability. However, implementing a cost-effective PCR remains challenging for rapid preventive medical action to the widespread pandemic diseases due to the absence of highly efficient and low-cost PCR chip-based POC molecular diagnostics. Here, this work reports an ultrafast metaphotonic PCR chip as a solution of a cost-effective and low-power-consumption POC device for the emerging global challenge of sustainable healthcare. This work designs a near-perfect photonic meta-absorber using ring-shaped titanium nitride to maximize the photothermal effect and realize rapid heating and cooling cycles during the PCR process. This work fabricates a large-area photonic meta-absorber on a 6-inch wafer cost-effectively using simple colloidal lithography. In addition, this work demonstrates 30 thermocycles from 65 (annealing temperature) to 95 °C (denaturation temperature) within 3 min 15 s, achieving an average 16.66 °C s-1 heating rate and 7.77 °C s-1 cooling rate during thermocycling, succeeding rapid metaphotonic PCR. This work believes a metaphotonic PCR chip can be used to create a low-cost, ultrafast molecular diagnostic chip with a meta-absorber.


Subject(s)
Polymerase Chain Reaction , Polymerase Chain Reaction/instrumentation , Polymerase Chain Reaction/methods , Titanium/chemistry , Humans , Lab-On-A-Chip Devices , Equipment Design
5.
Front Cell Infect Microbiol ; 14: 1419949, 2024.
Article in English | MEDLINE | ID: mdl-39119294

ABSTRACT

Human respiratory syncytial virus (HRSV) is the most prevalent pathogen contributing to acute respiratory tract infections (ARTI) in infants and young children and can lead to significant financial and medical costs. Here, we developed a simultaneous, dual-gene and ultrasensitive detection system for typing HRSV within 60 minutes that needs only minimum laboratory support. Briefly, multiplex integrating reverse transcription-recombinase polymerase amplification (RT-RPA) was performed with viral RNA extracted from nasopharyngeal swabs as a template for the amplification of the specific regions of subtypes A (HRSVA) and B (HRSVB) of HRSV. Next, the Pyrococcus furiosus Argonaute (PfAgo) protein utilizes small 5'-phosphorylated DNA guides to cleave target sequences and produce fluorophore signals (FAM and ROX). Compared with the traditional gold standard (RT-qPCR) and direct immunofluorescence assay (DFA), this method has the additional advantages of easy operation, efficiency and sensitivity, with a limit of detection (LOD) of 1 copy/µL. In terms of clinical sample validation, the diagnostic accuracy of the method for determining the HRSVA and HRSVB infection was greater than 95%. This technique provides a reliable point-of-care (POC) testing for the diagnosis of HRSV-induced ARTI in children and for outbreak management, especially in resource-limited settings.


Subject(s)
RNA, Viral , Respiratory Syncytial Virus Infections , Respiratory Syncytial Virus, Human , Sensitivity and Specificity , Humans , Respiratory Syncytial Virus, Human/genetics , Respiratory Syncytial Virus, Human/isolation & purification , Respiratory Syncytial Virus Infections/diagnosis , Respiratory Syncytial Virus Infections/virology , RNA, Viral/genetics , Infant , Pyrococcus furiosus/genetics , Pyrococcus furiosus/isolation & purification , Argonaute Proteins/genetics , Molecular Diagnostic Techniques/methods , Nucleic Acid Amplification Techniques/methods , Limit of Detection , Nasopharynx/virology , Respiratory Tract Infections/diagnosis , Respiratory Tract Infections/virology , Child, Preschool
6.
Talanta ; 279: 126566, 2024 Nov 01.
Article in English | MEDLINE | ID: mdl-39047627

ABSTRACT

Glutathione (GSH) is a non-protein tripeptide thiol that plays a prominent role in oxidative stress defense. GSH concentration is particularly critical in the neonatal period, especially for premature newborns that face increased susceptibility to oxidative stress. Monitoring GSH levels provides valuable insights into newborn health, helping to tailor care to their specific needs. The aim of this study was the development of a sensor specifically targeted for its use in neonatology, enabling GSH determination in only 2 µL of whole blood. The newly developed sensing system simplifies sample processing, addressing a critical need in clinical applications. Unlike current methods that demand fast pre-processing of relatively large sample volumes, expensive equipment, and skilled personnel, the developed approach streamlines the analytical process. By using 2 µL of whole blood, a single syringe filter for sample treatment, a deuterated internal standard (IS) for signal normalization, and Surface Enhanced Raman Scattering (SERS) spectroscopy with a silver colloid substrate for GSH detection, the set-up's characteristics are compatible with point-of-care applications. The analytical procedure was validated and applied to diverse populations including healthy adults (N = 63) and newborns (N = 35), yielding GSH concentration values ranging from 0.6 to 1.8 and 0.8-2.1 mM, respectively. This new optical sensor offers a quick and cost-effective solution to support the assessment of GSH levels in newborns that can greatly benefit not only neonatal care, but also the study of adult populations for health monitoring.


Subject(s)
Glutathione , Point-of-Care Systems , Spectrum Analysis, Raman , Humans , Glutathione/blood , Spectrum Analysis, Raman/methods , Infant, Newborn , Adult
7.
J Fungi (Basel) ; 10(7)2024 Jun 27.
Article in English | MEDLINE | ID: mdl-39057334

ABSTRACT

We present a case of a man immunocompromised due to myelodysplastic syndrome with Candida krusei fungemia who had a rising cell-free DNA (cfDNA) giant magnetoresistance (GMR) signal when tested daily using plasma blood samples. With the rise in GMR signal paralleling the development of skin lesions in this patient, we conclude that cfDNA can be used to indicate uncontrolled infection and thus help monitor response to therapy. This index patient provides evidence that an invasive fungal infection requires both direct antifungal therapy and an intact immune system to control the infection. This biosensing platform has been simplified to potentially serve as a point-of-care test, setting it apart by overcoming the three common barriers of cfDNA testing: complexity, cost, and time.

8.
Appl Microbiol Biotechnol ; 108(1): 414, 2024 Jul 10.
Article in English | MEDLINE | ID: mdl-38985204

ABSTRACT

Airborne animal viral pathogens can rapidly spread and become a global threat, resulting in substantial socioeconomic and health consequences. To prevent and control potential epidemic outbreaks, accurate, fast, and affordable point-of-care (POC) tests are essential. As a proof-of-concept, we have developed a molecular system based on the loop-mediated isothermal amplification (LAMP) technique for avian metapneumovirus (aMPV) detection, an airborne communicable agent mainly infecting turkeys and chickens. For this purpose, a colorimetric system was obtained by coupling the LAMP technique with specific DNA-functionalized AuNPs (gold nanoparticles). The system was validated using 50 different samples (pharyngeal swabs and tracheal tissue) collected from aMPV-infected and non-infected chickens and turkeys. Viral detection can be achieved in about 60 min with the naked eye, with 100% specificity and 87.88% sensitivity for aMPV. In summary, this novel molecular detection system allows suitable virus testing in the field, with accuracy and limit of detection (LOD) values highly close to qRT-PCR-based diagnosis. Furthermore, this system can be easily scalable to a platform for the detection of other viruses, addressing the current gap in the availability of POC tests for viral detection in poultry farming. KEY POINTS: •aMPV diagnosis using RT-LAMP is achieved with high sensitivity and specificity. •Fifty field samples have been visualized using DNA-nanoprobe validation. •The developed system is a reliable, fast, and cost-effective option for POCT.


Subject(s)
Chickens , Gold , Metapneumovirus , Molecular Diagnostic Techniques , Nucleic Acid Amplification Techniques , Paramyxoviridae Infections , Poultry Diseases , Sensitivity and Specificity , Metapneumovirus/genetics , Metapneumovirus/isolation & purification , Animals , Nucleic Acid Amplification Techniques/methods , Nucleic Acid Amplification Techniques/economics , Chickens/virology , Molecular Diagnostic Techniques/methods , Molecular Diagnostic Techniques/economics , Paramyxoviridae Infections/diagnosis , Paramyxoviridae Infections/veterinary , Paramyxoviridae Infections/virology , Poultry Diseases/virology , Poultry Diseases/diagnosis , Gold/chemistry , Turkeys , Metal Nanoparticles/chemistry , Limit of Detection , Colorimetry/methods , DNA, Viral/genetics
9.
medRxiv ; 2024 Jul 05.
Article in English | MEDLINE | ID: mdl-39006424

ABSTRACT

Diagnostic approaches that combine the high sensitivity and specificity of laboratory-based digital detection with the ease of use and affordability of point-of-care (POC) technologies could revolutionize disease diagnostics. This is especially true in infectious disease diagnostics, where rapid and accurate pathogen detection is critical to curbing the spread of disease. We have pioneered an innovative label-free digital detection platform that utilizes Interferometric Reflectance Imaging Sensor (IRIS) technology. IRIS leverages light interference from an optically transparent thin film, eliminating the need for complex optical resonances to enhance the signal by harnessing light interference and the power of signal averaging in shot-noise-limited operation to achieve virtually unlimited sensitivity. In our latest work, we have further improved our previous 'Single-Particle' IRIS (SP-IRIS) technology by allowing the construction of the optical signature of target nanoparticles (whole virus) from a single image. This new platform, 'Pixel-Diversity' IRIS (PD-IRIS), eliminated the need for z-scan acquisition, required in SP-IRIS, a time-consuming and expensive process, and made our technology more applicable to POC settings. Using PD-IRIS, we quantitatively detected the Monkeypox virus (MPXV), the etiological agent for Monkeypox (Mpox) infection. MPXV was captured by anti-A29 monoclonal antibody (mAb 69-126-3) on Protein G spots on the sensor chips and were detected at a limit-of-detection (LOD) - of 200 PFU/ml (~3.3 attomolar). PD-IRIS was superior to the laboratory-based ELISA (LOD - 1800 PFU/mL) used as a comparator. The specificity of PD-IRIS in MPXV detection was demonstrated using Herpes simplex virus, type 1 (HSV-1), and Cowpox virus (CPXV). This work establishes the effectiveness of PD-IRIS and opens possibilities for its advancement in clinical diagnostics of Mpox at POC. Moreover, PD-IRIS is a modular technology that can be adapted for the multiplex detection of pathogens for which high-affinity ligands are available that can bind their surface antigens to capture them on the sensor surface.

10.
Adv Sci (Weinh) ; 11(31): e2401386, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38894575

ABSTRACT

Since two-dimensionalal (2D) materials have distinct chemical and physical properties, they are widely used in various sectors of modern technologies. In the domain of diagnostic biodevices, particularly for point-of-care (PoC) biomedical diagnostics, 2D-based field-effect transistor biosensors (bio-FETs) demonstrate substantial potential. Here, in this review article, the operational mechanisms and detection capabilities of biosensing devices utilizing graphene, transition metal dichalcogenides (TMDCs), black phosphorus, and other 2D materials are addressed in detail. The incorporation of these materials into FET-based biosensors offers significant advantages, including low detection limits (LOD), real-time monitoring, label-free diagnosis, and exceptional selectivity. The review also highlights the diverse applications of these biosensors, ranging from conventional to wearable devices, underscoring the versatility of 2D material-based FET devices. Additionally, the review provides a comprehensive assessment of the limitations and challenges faced by these devices, along with insights into future prospects and advancements. Notably, a detailed comparison of FET-based biosensors is tabulated along with various other biosensing platforms and their working mechanisms. Ultimately, this review aims to stimulate further research and innovation in this field while educating the scientific community about the latest advancements in 2D materials-based biosensors.


Subject(s)
Biosensing Techniques , Point-of-Care Systems , Biosensing Techniques/methods , Biosensing Techniques/instrumentation , Humans , Transistors, Electronic , Graphite/chemistry , Equipment Design
11.
Diagnostics (Basel) ; 14(11)2024 May 24.
Article in English | MEDLINE | ID: mdl-38893615

ABSTRACT

Saliva has shown considerable promise as a diagnostic medium for point-of-care (POC) and over-the-counter (OTC) diagnostic devices due to the non-invasive nature of its collection. However, a significant limitation of saliva-based detection is undesirable interference in a sensor's readout caused by interfering components in saliva. In this study, we develop standardized sample treatment procedures to eliminate bubbles and interfering molecules while preserving the sample's target molecules such as spike (S) protein and glucose. We then test the compatibility of the pretreatment system with our previously designed SARS-CoV-2 and glucose diagnostic biosensing systems for detecting S protein and glucose in subject saliva. Ultimately, the effectiveness of each filter in enhancing biomarker sensitivity is assessed. The results show that a 20 mg nylon wool (NW) filter shows an 80% change in viscosity reduction with only a 6% reduction in protein content, making it an appropriate filter for the salivary S protein diagnostic system. Meanwhile, a 30 mg cotton wool (CW) filter is identified as the optimal choice for salivary glucose detection, achieving a 90% change in viscosity reduction and a 60.7% reduction in protein content with a minimal 4.3% reduction in glucose content. The NW pretreatment filtration significantly improves the limit of detection (LOD) for salivary S protein detection by five times (from 0.5 nM to 0.1 nM) and it reduces the relative standard deviation (RSD) two times compared to unfiltered saliva. Conversely, the CW filter used for salivary glucose detection demonstrated improved linearity with an R2 of 0.99 and a sensitivity of 36.6 µA/mM·cm2, over twice as high as unfiltered saliva. This unique filtration process can be extended to any POC diagnostic system and optimized for any biomarker detection, making electrochemical POC diagnostics more viable in the current market.

12.
Diagnostics (Basel) ; 14(11)2024 May 26.
Article in English | MEDLINE | ID: mdl-38893633

ABSTRACT

In April 2020, the Aboriginal and Torres Strait Islander COVID-19 Point-of-Care (POC) Testing Program was initiated to improve access to rapid molecular-based SARS-CoV-2 detection in First Nations communities. At capacity, the program reached 105 health services across Australia. An external review estimated the program contributed to averting between 23,000 and 122,000 COVID-19 infections within 40 days of the first infection in a remote community, equating to cost savings of between AU$337 million and AU$1.8 billion. Essential to the quality management of this program, a customised External Quality Assessment (EQA) program was developed with the Royal College of Pathologists of Australasia Quality Assurance Programs (RCPAQAP). From July 2020 to May 2022, SARS-CoV-2 EQA participation ranged from 93 to 100%. Overall concordance of valid EQA results was high (98%), with improved performance following the first survey. These results are consistent with those reported by 12 Australian and 4 New Zealand laboratories for three SARS-CoV-2 RNA EQA surveys in March 2020, demonstrating that SARS-CoV-2 RNA POC testing in primary care settings can be performed to an equivalent laboratory analytical standard. More broadly, this study highlights the value of quality management practices in real-world testing environments and the benefits of ongoing EQA program participation.

13.
Entropy (Basel) ; 26(4)2024 Mar 26.
Article in English | MEDLINE | ID: mdl-38667838

ABSTRACT

Recently, with more portable diagnostic devices being moved to people anywhere, point-of-care (PoC) imaging has become more convenient and more popular than the traditional "bed imaging". Instant image segmentation, as an important technology of computer vision, is receiving more and more attention in PoC diagnosis. However, the image distortion caused by image preprocessing and the low resolution of medical images extracted by PoC devices are urgent problems that need to be solved. Moreover, more efficient feature representation is necessary in the design of instant image segmentation. In this paper, a new feature representation considering the relationships among local features with minimal parameters and a lower computational complexity is proposed. Since a feature window sliding along a diagonal can capture more pluralistic features, a Diagonal-Axial Multi-Layer Perceptron is designed to obtain the global correlation among local features for a more comprehensive feature representation. Additionally, a new multi-scale feature fusion is proposed to integrate nonlinear features with linear ones to obtain a more precise feature representation. Richer features are figured out. In order to improve the generalization of the models, a dynamic residual spatial pyramid pooling based on various receptive fields is constructed according to different sizes of images, which alleviates the influence of image distortion. The experimental results show that the proposed strategy has better performance on instant image segmentation. Notably, it yields an average improvement of 1.31% in Dice than existing strategies on the BUSI, ISIC2018 and MoNuSeg datasets.

14.
Transfusion ; 64(5): 881-892, 2024 May.
Article in English | MEDLINE | ID: mdl-38591151

ABSTRACT

BACKGROUND: A life-threatening anaphylactic shock can occur if a patient with undiagnosed immunoglobulin A (IgA) deficiency (i.e., IgA levels <500 ng/mL) receives IgA-containing blood, hence the need for a rapid, point-of-care (POC) method for IgA deficiency screening. Enzyme-linked immunosorbent assay (ELISA) is routinely used to detect IgA, but this method requires trained specialists and ≥24 h to obtain a result. We developed a surface plasmon resonance (SPR)-based protocol to identify IgA-deficient patients or donors within 1 h. MATERIALS AND METHODS: The SPR sensor relies on the detection of IgAs captured by primary antibodies adsorbed on the SPR chip and quantified with secondary antibodies. The sensor was calibrated from 0 to 2000 ng/mL in buffer, IgA-depleted human serum, and plasma samples from IgA-deficient individuals. A critical concentration of 500 ng/mL was set for IgA deficiency. The optimized sensor was then tested on eight plasma samples with known IgA status (determined by ELISA), including five with IgA deficiency and three with normal IgA levels. RESULTS: The limit of detection was estimated at 30 ng/mL in buffer and 400 ng/mL in diluted plasma. The results obtained fully agreed with ELISA among the eight plasma samples tested. The protocol distinguished IgA-deficient from normal samples, even for samples with an IgA concentration closer to critical concentration. DISCUSSION: In conclusion, we developed a reliable POC assay for the quantification of IgA in plasma. This test may permit POC testing at blood drives and centralized centers to maintain reserves of IgA-deficient blood and in-hospital testing of blood recipients.


Subject(s)
IgA Deficiency , Immunoglobulin A , Surface Plasmon Resonance , Humans , Surface Plasmon Resonance/methods , Surface Plasmon Resonance/instrumentation , Immunoglobulin A/blood , IgA Deficiency/blood , IgA Deficiency/diagnosis , Enzyme-Linked Immunosorbent Assay/methods
15.
Front Med Technol ; 6: 1360510, 2024.
Article in English | MEDLINE | ID: mdl-38425422

ABSTRACT

Herein, advancements in electroanalytical devices for the simultaneous detection of diverse breast cancer (BC) markers are demonstrated. This article identifies several important areas of exploration for electrochemical diagnostics and highlights important factors that are pivotal for the successful deployment of novel bioanalytical devices. We have highlighted that the limits of detection (LOD) reported for the multiplex electrochemical biosensor can surpass the sensitivity displayed by current clinical standards such as ELISA, FISH, and PCR. HER-2; a breast cancer marker characterised by increased metastatic potential, more aggressive development, and poor clinical outcomes; can be sensed with a LOD of 0.5 ng/ml using electrochemical multiplex platforms, which falls within the range of that measured by ELISA (from picogram/ml to nanogram/ml). Electrochemical multiplex biosensors are reported with detection limits of 0.53 ng/ml and 0.21 U/ml for MUC-1 and CA 15-3, respectively, or 5.8 × 10-3 U/ml for CA 15-3 alone. The sensitivity of electrochemical assays is improved when compared to conventional analysis of MUC-1 protein which is detected at 11-12 ng/ml, and ≤30 U/ml for CA 15-3 in the current clinical blood tests. The LOD for micro-ribonucleic acid (miRNA) biomarkers analyzed by electrochemical multiplex assays were all notedly superior at 9.79 × 10-16 M, 3.58 × 10-15 M, and 2.54 × 10-16 M for miRNA-155, miRNA-21, and miRNA-16, respectively. The dogma in miRNA testing is the qRT-PCR method, which reports ranges in the ng/ml level for the same miRNAs. Breast cancer exosomes, which are being explored as a new frontier of biosensing, have been detected electrochemically with an LOD of 103-108 particles/mL and can exceed detection limits seen by the tracking and analysis of nanoparticles (∼ 107 particles/ml), flow cytometry, Western blotting and ELISA, etc. A range of concentration at 78-5,000 pg/ml for RANKL and 16-1,000 pg/ml for TNF is reported for ELISA assay while LOD values of 2.6 and 3.0 pg/ml for RANKL and TNF, respectively, are demonstrated by the electrochemical dual immunoassay platform. Finally, EGFR and VEGF markers can be quantified at much lower concentrations (0.01 and 0.005 pg/ml for EGFR and VEGF, respectively) as compared to their ELISA assays (EGRF at 0.31-20 ng/ml and VEGF at 31.3-2,000 pg/ml). In this study we hope to answer several questions: (1) Are the limits of detection (LODs) reported for multiplex electrochemical biosensors of clinical relevance and how do they compare to well-established methods like ELISA, FISH, or PCR? (2) Can a single sensor electrode be used for the detection of multiple markers from one blood drop? (3) What mechanism of electrochemical biosensing is the most promising, and what technological advancements are needed to utilize these devices for multiplex POC detection? (4) Can nanotechnology advance the sensitive and selective diagnostics of multiple BC biomarkers? (5) Are there preferred receptors (antibody, nucleic acid or their combinations) and preferred biosensor designs (complementary methods, sandwich-type protocols, antibody/aptamer concept, label-free protocol)? (6) Why are we still without FDA-approved electrochemical multiplex devices for BC screening?

16.
Anal Sci ; 40(6): 1101-1110, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38468109

ABSTRACT

Discovering the balance between toxicity and efficacy for many drugs requires therapeutic drug monitoring (TDM) of their concentrations in the blood. Here, a hot-embossed microfluidic device with a new design integrated to a nanofracture is presented for purification of blood samples from numerous proteins and cells, allowing to the separation of small molecules from blood matrix. The device was used to separate and quantitatively detect tamsulosin drug after derivatization with fluorescamine reagent, allowing converting it from a neutral molecule into a charged fluorescent complex under the experimental conditions, and thus its separation by electrophoresis. The device is portable and easy operated, and the presented method showed good linearity (R2 = 0.9948) over a concentration range of 0.1-1 µg/mL. The relative standard deviation (RSD%) was below 10% (n = 3), indicating good precisions, and the limit of detection (LOD) and limit of quantitation (LOQ) values were estimated to be 0.1 and 0.55 µg/mL, respectively. Whole blood samples from 10 patients with benign prostatic hyperplasia (BPH) were analyzed, showing good percentage recoveries of tamsulosin in whole blood. This point-of-care (POC), low-cost method could increase the convenience of patients and doctors, make therapies safer, and make TDM available in different regions and places.


Subject(s)
Drug Monitoring , Point-of-Care Systems , Prostatic Hyperplasia , Tamsulosin , Tamsulosin/blood , Humans , Prostatic Hyperplasia/blood , Prostatic Hyperplasia/drug therapy , Drug Monitoring/instrumentation , Drug Monitoring/methods , Male , Nanotechnology , Lab-On-A-Chip Devices
17.
Biosens Bioelectron ; 251: 116097, 2024 May 01.
Article in English | MEDLINE | ID: mdl-38330774

ABSTRACT

Severe periodontitis affects nearly 1 billion individuals worldwide, highlighting the need for early diagnosis. Here, an integrated system consisting of a microfluidic chip and a portable point-of-care (POC) diagnostic device is developed using a polymethyl methacrylate (PMMA) chip fabrication and a three-dimensional printing technique, which is automatically controlled by a custom-designed smartphone application to routinely assess the presence of a specific periodontitis biomarker, odontogenic ameloblast-associated protein (ODAM). A sandwich-type fluorescence aptasensor is developed on a microfluidic chip, utilizing aptamer pair (MB@OD64 and OD35@FAM) selectively binding to target ODAM. Then this microfluidic chip is integrated into an automated Internet of Things (IoT)-based POC device, where fluorescence intensity, as a signal, from the secondary aptamer binding to ODAM in a sandwich-type binding reaction on the microfluidic chip is measured by a complementary metal oxide semiconductor (CMOS) camera with a 488 nm light-emitting diode (LED) excitation source. Obtained signals are processed by a microprocessor and visualized on a wirelessly connected smartphone application. This integrated biosensor system allows the rapid and accurate detection of ODAM within 30 min with a remarkable limit of detection (LOD) of 0.011 nM under buffer conditions. Clinical application is demonstrated by successfully distinguishing between low-risk and high-risk individuals with 100 % specificity. A strong potential in the translation of this fluorescence-based microfluidic aptasensor integrated with an IoT-based POC system is expected to be employed for non-invasive, on-site, rapid, and accurate ODAM detection, facilitating periodontitis diagnosis.


Subject(s)
Biosensing Techniques , Internet of Things , Periodontal Diseases , Periodontitis , Humans , Periodontal Diseases/diagnosis , Periodontitis/metabolism , Proteins
18.
Talanta ; 271: 125628, 2024 May 01.
Article in English | MEDLINE | ID: mdl-38219320

ABSTRACT

This article presents the development of a photothermal biosensing integrated with microfluidic paper-based analytical device (PT-µPAD) as a quantitative biosensor method for monitoring sarcosine in human control urine, plasma, and serum samples. The device utilizes gold nanoparticles (AuNPs) as both a peroxidase-like nanozyme and a photothermal substrate to enable sarcosine detection. In our PT-µPAD, hydrogen peroxide (H2O2) is generated through the oxidation of sarcosine by a sarcosine oxidase (SOx) enzyme. Subsequently, the H2O2 flows through the paper microchannels to the detection zone, where it etches the pre-deposited AuNPs, inducing a temperature change upon exposure by a 532 nm laser. The temperature variation is then measured using a portable and inexpensive infrared thermometer. Under optimized conditions, we obtained a linear range between 10.0 and 40.0 nmol L-1 (R2 = 0.9954) and a detection limit (LOD) of 32.0 pmol L-1. These values fall within the clinical range for sarcosine monitoring in prostate cancer diagnostics in humans. Moreover, our approach exhibits high selectivity without interfering effects. Recovery studies in various human control samples demonstrated a range of 99.05-102.11 % with the highest RSD of 2.25 %. The PT-µPAD was further validated for sarcosine determination in human control urine and compared with a commercial ELISA assay, revealing no significant difference between these two methods at a 95 % confidence level. Overall, our proposed sarcosine biosensor is well-suited for prostate cancer monitoring, given its affordability, sensitivity, and user-friendliness, even for unskilled individuals. Moreover, this strategy has promising prospects for broader applications, potentially detecting various biomarkers as a point-of-care (POC) diagnostic tool.


Subject(s)
Biosensing Techniques , Metal Nanoparticles , Prostatic Neoplasms , Male , Humans , Sarcosine/analysis , Gold , Microfluidics , Hydrogen Peroxide , Prostatic Neoplasms/diagnosis , Biosensing Techniques/methods
19.
ACS Sens ; 8(12): 4442-4467, 2023 Dec 22.
Article in English | MEDLINE | ID: mdl-38091479

ABSTRACT

In recent years, there has been a notable increase in interest surrounding nanozymes due to their ability to imitate the functions and address the limitations of natural enzymes. The scientific community has been greatly intrigued by the study of nanoceria, primarily because of their distinctive physicochemical characteristics, which include a variety of enzyme-like activities, affordability, exceptional stability, and the ability to easily modify their surfaces. Consequently, nanoceria have found extensive use in various biosensing applications. However, the impact of its redox activity on the enzymatic catalytic mechanism remains a subject of debate, as conflicting findings in the literature have presented both pro-oxidant and antioxidant effects. Herein, we creatively propose a seesaw model to clarify the regulatory mechanism on redox balance and survey possible mechanisms of multienzyme mimetic properties of nanoceria. In addition, this review aims to showcase the latest advancements in this field by systematically discussing over 180 research articles elucidating the significance of ceria-based nanozymes in enhancing, downsizing, and enhancing the efficacy of point-of-care (POC) diagnostics. These advancements align with the ASSURED criteria established by the World Health Organization (WHO). Furthermore, this review also examines potential constraints in order to offer readers a concise overview of the emerging role of nanoceria in the advancement of POC diagnostic systems for future biosensing applications.


Subject(s)
Cerium , Point-of-Care Systems , Oxidation-Reduction , Cerium/chemistry , Antioxidants
20.
Microbiol Spectr ; 11(6): e0277223, 2023 Dec 12.
Article in English | MEDLINE | ID: mdl-37962351

ABSTRACT

IMPORTANCE: In hospitals during the COVID-19 pandemic, laboratory testing was important to reduce SARS-CoV-2 transmissions, while facilitating patient flow in the emergency department and pre-operative settings, and allowing for the safe return to work of exposed healthcare workers. Delayed test results from laboratory nucleic acid amplification tests (NAATs) posed a barrier to maximizing efficient patient flow and minimizing staffing shortages. This quality improvement project sought to evaluate the analytical and clinical performance of the Lucira Check-It COVID-19 Test, a point-of-care test that used NAAT technology, in the perioperative setting, emergency department, and community testing sites. We found the Lucira Check-It to have comparable performance to laboratory NAATs. It can be employed with little training for specimen collection, processing, and interpretation, and at a cost justifiable from the resources saved from avoiding sample transport and laboratory testing.


Subject(s)
COVID-19 , Humans , COVID-19/diagnosis , SARS-CoV-2/genetics , Clinical Laboratory Techniques/methods , COVID-19 Testing , Molecular Diagnostic Techniques/methods , Pandemics , Sensitivity and Specificity
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