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Nowadays, kidney dysfunction is a common health issue due to the modernized lifestyle. Even though medications are commercially available to treat kidney diseases, early diagnosis is crucial and challenging. Clinically, measuring urine creatinine and pH has gained significant interest as a way to diagnose kidney diseases early. In the present work, we attempted to develop a low-cost, robust, accurate and naked-eye colorimetric method to determine both creatinine levels and pH variations in artificial urine samples using a simple 3D-printed hybrid microfluidic device. Creatinine was detected by the incorporation of the traditional Jaffe test onto the hybrid paper-PMMA microfluidic device and pH (4-8) was measured by a simple anthocyanin test. Notably, the tests were established without employing any sophisticated or costly instrument clusters. The developed 3D-printed microfluidic probe showed a limit of detection (LOD) of 0.04 mM for creatinine over a concentration range of 1-10 mM, with a regression coefficient ( R 2 ) of 0.995 in laboratory conditions. Interestingly, the experimental data obtained with artificial urine exhibited a wide linear range from 0.1 mM to 5 mM under different pH values ranging from 4 to 8 in the presence of matrices commonly found in urine samples other than proteins, indicating the potential use of this method in pre-clinical analysis. Since the wide linear range of urine creatinine in artificial urine samples falls well below the clinically relevant concentrations in humans (0.07-0.27 mM), the developed lab-on-chip device is further suitable for clinical evaluation with proper ethical clearance. This 3D-printed hybrid microfluidic colorimetry-based creatinine detection and pH indicator platform can be beneficial in the healthcare sector due to the on-site testing capability, cost-effectiveness, ease of use, robustness, and instrument-free approach.

This study introduces the development of a photothermal-based microfluidic paper analytical device (PT-µPAD) integrated with near-field communication (NFC) technology and smartphone readout for enzyme-free glucose quantification in human samples. With the properties of gold nanoparticles (AuNPs) both as a nanozyme and as a photothermal substrate, there is no need for costly reagents like enzymes or a readout instrumentation for the selective and sensitive detection of glucose. In PT-µPADs, AuNPs are etched by hydrogen peroxide (H 2 O 2 ) generated from glucose catalysis. Photothermal detection from the plasmonic heating of these AuNPs when illuminated by a 533nm LED light source is achieved by inserting the PT-µPAD sensor into a portable NFC platform suitable for smartphone readout. Temperature variation is directly proportional to the glucose concentration. After optimization, we acquired a linear range between 5.0 and 20.0 µmol L -1 ( R 2 = 0.9967) and a limit of detection (LOD) of 25.0 nmol L -1 for glucose. Additionally, while our sensor does not utilize any enzyme, it is remarkably selective to glucose with no effects from interferences. Recovery studies in various human control samples indicated a range of 99.73-102.66% with the highest RSD of 3.53%, making it highly accurate and precise. Moreover, our method is more sensitive than other methods relying on conventional µPADs for glucose sensing. By integrating the potential benefits of microfluidics, nanomaterials as nanozymes, and NFC technology for wireless readout, our sensor demonstrates great promise as an accessible, affordable, and shelf-stable device for glucose quantification. Moreover, this concept can be extended to detect other molecules of interest as a point-of-care (POC) diagnostics device.

We reported a colorimetric paper-based device by integrating the modified acid RNA-cleaving DNAzymes (MaRCD-EC1) for highly sensitive (detection limit = 10 2 CFU mL -1 ), and rapid (within 30 min) detection of E. coli without amplification. This device exhibited a clinical sensitivity of 100% and a specificity of 100% in identifying E. coli -associated urinary tract infections (UTIs) using the clinical urine samples.

Safe, accurate, and reliable analysis of urinary biomarkers is clinically important for early detection and monitoring of the progression of chronic kidney disease (CKD), as it has become one of the world's most prevalent non-communicable diseases. However, current technologies for measuring urinary biomarkers are either time-consuming and limited to well-equipped hospitals or lack the necessary sensitivity for quantitative analysis and post a health risk to frontline practitioners. Here we report a robust paper-based dual functional biosensor, which is integrated with the clinical urine sampling vial, for the simultaneous and quantitative analysis of pH and glucose in urine. The pH sensor was fabricated by electrochemically depositing IrOx onto a paper substrate using optimised parameters, which enabled an ultrahigh sensitivity of 71.58 mV pH -1 . Glucose oxidase (GOx) was used in combination with an electrochemically deposited Prussian blue layer for the detection of glucose, and its performance was enhanced by gold nanoparticles (AuNPs), chitosan, and graphite composites, achieving a sensitivity of 1.5 μA mM -1 . This dual function biosensor was validated using clinical urine samples, where a correlation coefficient of 0.96 for pH and 0.98 for glucose detection was achieved with commercial methods as references. More importantly, the urine sampling vial was kept sealed throughout the sample-to-result process, which minimised the health risk to frontline practitioners and simplified the diagnostic procedures. This diagnostic platform, therefore, holds high promise as a rapid, accurate, safe, and user-friendly point-of-care (POC) technology for the analysis of urinary biomarkers in frontline clinical settings.

A new analytical method for chronic kidney disease (CKD) detection utilizing paper spray mass spectrometry (PS-MS) combined with machine learning is presented. The analytical protocol is rapid and simple, based on metabolic profile alterations in urine. Anonymized raw urine samples were deposited (10 μL each) onto pointed PS-MS sample strips. Without waiting for the sample to dry, 75 μL of acetonitrile and high voltage were applied to the strips, using high resolution mass spectrometry measurement (15 s per sample) with polarity switching to detect a wide range of metabolites. Random forest machine learning was used to classify the resulting data. The diagnostic performance for the potential diagnosis of CKD was evaluated for accuracy, sensitivity, and specificity, achieving results >96% for the training data and >91% for validation and test data sets. Metabolites selected by the classification model as up- or down-regulated in healthy or CKD samples were tentatively identified and in agreement with previously reported literature. The potential utilization of this approach to discriminate albuminuria categories (normo, micro, and macroalbuminuria) was also demonstrated. This study indicates that PS-MS combined with machine learning has the potential to be used as a rapid and simple diagnostic tool for CKD.

Paper-based sensors offer an affordable yet powerful platform for field and point-of-care (POC) testing due to their self-pumping ability and utility for many different analytical measurements. When combined with electrochemical detection using small and portable electronics, sensitivity and selectivity of the paper devices can be improved over naked eye detection without sacrificing portability. Herein, we review how the field of electrochemical paper-based analytical devices (ePADs) has grown since it was introduced a decade ago. We start by reviewing fabrication methods relevant to ePADs with more focus given to the electrode fabrication, which is fundamental for electrochemical sensing. Multiple sensing approaches applicable to ePADs are then discussed and evaluated to present applicability, advantages and challenges associated with each approach. Recent applications of ePADs in the fields of clinical diagnostics, environmental testing, and food analysis are also presented. Finally, we discuss how the current ePAD technologies have progressed to meet the analytical and practical specifications required for field and/or POC applications, as well as challenges and outlook. [ABSTRACT FROM AUTHOR]

A simple, rapid chemical coating and patterning method was developed and optimized for paper-based substrates for use in paper spray mass spectrometry (PS-MS). A variety of chlorosilanes were explored for coating paper substrates, and their effectiveness in forming hydrophobic surfaces was characterized via contact angle goniometry, scanning electron microscopy, and energy dispersive X-ray spectroscopy. Trichloromethylsilane was selected as the primary coating agent because of the short time required to produce a hydrophobic surface (contact angle > 130°), as well as the ease of patterning. Patterning was performed using 3D-printed masks and an oxygen/plasma cleaner. Optimal mask thickness and oxygen/plasma cleaning parameters were determined to produce channels varying from 0.5 to 2.5 mm in width. The effectiveness of the patterned substrates for PS-MS was determined via analysis of four antiretrovirals: emtricitabine, lamivudine, efavirenz, and dolutegravir. Calibration curves were made for each antiretroviral at varying channel widths, and the limits of detection and limits of quantification for each drug were determined. These results show that this patterning method results in an average 7.2-fold improvement in sensitivity and an average 190-fold improvement in limits of detection over uncoated paper substrates in a neat matrix. In a proof-of-concept experiment, calibration curves were generated for each antiretroviral in urine. A patterned paper substrate with a 2-mm channel resulted in an average 7.4-fold improvement in sensitivity and an average 18-fold improvement in limits of detection over uncoated paper substrates.

The correct interpretation of the result from a point-of-care device is crucial for an accurate and rapid diagnosis to guide subsequent treatment. Lateral flow tests (LFTs) use a well-established format that was designed to simplify the user experience. However, the LFT device architecture is inherently limited to detecting analytes that can be captured by molecular recognition. Microfluidic paper-based analytical devices (μPADs), like LFTs, have the potential to be used in diagnostic applications at the point of care. However, μPADs have not gained significant traction outside of academic laboratories, in part, because they have often demonstrated a lack of homogeneous shape or color in signal outputs, which consequently can lead to inaccurate interpretation of results by users. Here, we demonstrate a new class of μPADs that form colorimetric signals at the interfaces of converging liquid fronts ( i.e. , lines) to control where colorimetric signals are formed without relying on capture techniques. We demonstrate our approach by developing assays for three classes of analytes-an ion, an enzyme, and a small molecule-to measure using iron(III), acetylcholinesterase, and lactate, respectively. Additionally, we show these devices have the potential to support multiplexed assays by generating multiple lines in a common readout zone. These results highlight the ability of this new paper-based device architecture to aid the interpretation of assays that create soluble products by using flow to constrain those colorimetric products in a familiar, line-format output.

Lab-on-a-paper-based devices are promising alternatives to the existing arduous techniques for point-of-need monitoring. The present work reports an instant and facile method to produce a microfluidic paper-based analytical device (μPAD). The fabricated μPAD has been used to detect hypochlorite (OCl - ) by incorporating newly synthesized chromo-fluorogenic ratiometric probes 1 and 2 into the sample reception zone. The probes showed high selectivity and fast response (<10 s) toward OCl - with an excellent linear relationship in the concentration range of 0-100 μM. The concentration-dependent fluorometric change driven by the reaction of 1@μPAD with OCl - has been monitored using gel-doc imaging systems, which is unprecedented. Digitizing the intensity of the colour solution with different mathematical models of colour has developed a straightforward method for monitoring OCl - without any interference from its competitors. 1@μPAD can detect OCl - at ∼10 times lower than the WHO recommended limit. The detection limit of 1@μPAD via a digital camera-based fluorescence technique was found to be better over digital camera-based cuvette assays. Therefore, 1@μPAD has been successfully utilized to monitor OCl - in actual environmental water samples with portability, ease of use, and sensitivity. The analytical RSD was found to be ≤3% based on fluorimetric detection using 1@μPAD. The chemodosimetric reaction between OCl - and the probe was evidenced by UV-vis and fluorescence spectroscopy, 1 H NMR, and ESI-MS. The rapid response time, biocompatibility, low cytotoxicity, 100% aqueous solubility, ratiometric feature, and exclusive OCl - selectivity over other competitive ROS/RNS successfully lead to the application of the probes for bioimaging of exogenous as well as endogenous OCl - in normal cells (HEK293) and cancerous cells (HeLa).

Paper-based analytical devices (e.g. lateral flow assays) are highly advantageous as portable diagnostic systems owing to their low costs and ease of use. Because of their low sensitivity and detection limits for biomolecules, these devices have several limitations in applications for real-field diagnosis. Here, we demonstrate a paper-based preconcentration enhanced lateral flow assay using a commercial β-hCG-based test. Utilizing a simple 9 V battery operation with a low power consumption of approximately 81 μW, we acquire a 25-fold preconcentration factor, demonstrating a clear sensitivity enhancement in the colorimetric lateral flow assay; consequently, clear colors are observed in a rapid kit test line, which cannot be monitored without preconcentration. This device can also facilitate a semi-quantitative platform using the saturation value and/or color intensity in both paper-based colorimetric assays and smartphone-based diagnostics. [ABSTRACT FROM AUTHOR]

This communication describes the development of polyvinyl chloride electrochemical system in which a paper layer loaded with reagents is inserted into the device, demonstrating a new concept of a paper card-like pad for a reagent-free and easy measurement of the target analyte in solution. This device detects glucose in artificial tears in the range of 0.2-2 mM with a detection limit of 50 μM by simply adding the artificial tears to the paper card-like pad. The novel configuration goes beyond the state of the art, widening the application range of paper in the design of smart analytical devices.

A scalable, and cost-effective method was employed to prepare self-supported CuSn bimetallic catalyst on carbon paper. The obtained CuSn catalyst demonstrates high faradaic efficiency of CO around or above 90% at a broad potential range from -0.7 to -1.8 V vs. reversible hydrogen electrode, greatly surpassing Cu or Sn counterparts.

The lack of sensitivity is a major problem among microfluidic paper-based analytical devices (μPADs) for early disease detection and diagnosis. Accordingly, the present study presents a method for improving the enrichment factor of low-concentration biomarkers by using shallow paper-based channels realized through a double-sided wax-printing process. In addition, the enrichment factor is further enhanced by exploiting the ion concentration polarization (ICP) effect on the cathodic side of the nanoporous membrane, in which a stationary sample plug is obtained. The occurrence of ICP on the shallow-channel μPAD is confirmed by measuring the current–voltage response as the external voltage is increased from 0 to 210 V (or the field strength from 0 to 1.05 × 104 V m−1) over 600 s. In addition, to the best of our knowledge, the electroosmotic flow (EOF) speed on the μPAD fabricated with a wax-channel is measured for the first time using a current monitoring method. The experimental results show that for a fluorescein sample, the concentration factor is increased from 130-fold in a conventional full-thickness paper channel to 944-fold in the proposed shallow channel. Furthermore, for a fluorescein isothiocyanate-labeled bovine serum albumin (FITC-BSA) sample, the proposed shallow-channel μPAD achieves an 835-fold improvement in the concentration factor. The concentration technique presented here provides a novel strategy for enhancing the detection sensitivity of μPAD applications. [ABSTRACT FROM AUTHOR]

Microfluidic paper-based analytical devices (microPADs) are emerging as simple-to-use, low-cost point-of-care testing platforms. Such devices are mostly fabricated at present by creating hydrophobic barriers using wax or photoresist patterning on porous paper sheets. Even though devices fabricated using these methods are used and tested with a wide variety of analytes, still they pose many serious practical limitations for low-cost automated mass fabrication for their widespread applicability. We present an affordable and simple two-step process - cut and heat (CH-microPADs) - for the selective fabrication of hydrophilic channels and reservoirs on a wide variety of porous media such as tissue/printing/filter paper and cloth types, such as cotton and polyester, by a lamination process. The technique presents many advantages as compared to existing commonly used methods. The devices possess excellent mechanical strength against bending, folding and twisting, making them virtually unbreakable. They are structurally flexible and show good chemical resistance to various solvents, acids and bases, presenting widespread applicability in areas such as clinical diagnostics, biological sensing applications, food processing, and the chemical industry. Fabricated paper media 96 well-plate CH-microPAD configurations were tested for cell culture applications using mice embryonic fibroblasts and detection of proteins and enzymes using ELISA. With a simple two-step process and minimal human intervention, the technique presents a promising step towards mass fabrication of inexpensive disposable diagnostic devices for both resource-limited and developed regions.

Microfluidic paper-based analytical devices (μPADs) have attracted great attention as potential candidates for point-of-care testing (POCT). Nevertheless, only a limited number of μPADs expected to satisfy the standard of Clinical Laboratory Improvement Amendments (CLIA) waived tests as issued by the US Food and Drug Administration (FDA) have been reported. This work introduces a "traffic light type μPAD", enabling highly intuitive semi-quantitative equipment-free naked-eye readout with no need for calibration, subjective interpretation or calculation. Assay results are displayed as traffic light colours reporting 5 analyte concentration levels (green/green & yellow/yellow/yellow & red/red). The device has been designed to never display all three colours simultaneously, eliminating any risk for misinterpretation. The mechanism relies on the modulation of sample flow through a network of paperfluidic channels modified with a hydrophobic to hydrophilic phase-switching substance responsive to H 2 O 2 . User operation is limited to sample application, followed by observing a clear and time-independent traffic light signal after approximately 10-30 min. Multiple factors influencing the H 2 O 2 concentration-dependent appearance of a specific traffic light signal were studied. Making use of the possibilities for customising the concentration threshold levels for traffic light colour appearance, quantification of glucose at 5 levels in a clinically relevant concentration range was demonstrated in artificial urine as a model proof-of-concept. This platform is expected to offer the possibility for the future detection of other important metabolites.

A new method for making superhydrophobic (SH) paper based on spraying a nanocomposite film is developed. Furthermore, manipulating the wetting characteristics of SH paper has been demonstrated through a new method, i.e.printing solid grey patterns of different intensities with simple printing technology (home or office grade ink-jet and laser printers). It has been found that for a range of ink intensities (0–85%), water drop mobility can be changed at a different rate (almost independently) from repellency. The repellency of water decreases minimally up to 85% ink intensity with a sharp decrease up to 100% ink intensity. Drop mobility remains constant up to 30% ink intensity with a steady decrease up to 100% ink intensity. It was observed that using ink-jet or laser printing would yield different results for the change of mobility or repellency with higher amounts of ink/toner used. Being able to achieve almost independent control of water drop mobility over water drop repellency on SH paper would allow inexpensive lab-on-paper devices to be used for sampling, mixing and transport of liquids. [ABSTRACT FROM AUTHOR]

We report a method for fabricating inexpensive microfluidic platforms on paper using laser treatment. Any paper with a hydrophobic surface coating (e.g., parchment paper, wax paper, palette paper) can be used for this purpose. We were able to selectively modify the surface structure and property (hydrophobic to hydrophilic) of several such papers using a CO2laser. We created patterns down to a minimum feature size of 62 ± 1 µm. The modified surface exhibited a highly porous structure which helped to trap/localize chemical and biological aqueous reagents for analysis. The treated surfaces were stable over time and were used to self-assemble arrays of aqueous droplets. Furthermore, we selectively deposited silica microparticles on patterned areas to allow lateral diffusion from one end of a channel to the other. Finally, we demonstrated the applicability of this platform to perform chemical reactions using luminol-based hemoglobin detection. [ABSTRACT FROM AUTHOR]

This paper describes the fabrication and the performance of microfluidic paper-based electrochemical sensing devices (we call the microfluidic paper-based electrochemical devices, µPEDs). The µPEDs comprise paper-based microfluidic channels patterned by photolithography or wax printing, and electrodes screen-printed from conducting inks (e.g., carbon or Ag/AgCl). We demonstrated that the µPEDs are capable of quantifying the concentrations of various analytes (e.g., heavy-metal ions and glucose) in aqueous solutions. This low-cost analytical device should be useful for applications in public health, environmental monitoring, and the developing world. [ABSTRACT FROM AUTHOR]

Paper has been proposed as an inexpensive and versatile carrier for microfluidics devices with abilities well beyond simple capillary action for pregnancy tests and the like. Unlike standard microfluidics devices, extracting a fluid from the paper is a challenge and a drawback to its broader use. Here, we extract fluid from narrow paper strips using surface acoustic wave (SAW) irradiation that subsequently atomizes the extracted fluid into a monodisperse aerosol for use in mass spectroscopy, medical diagnostics, and drug delivery applications. Two protein molecules, ovalbumin and bovine serum albumin (BSA), have been preserved in paper and then extracted using atomized mist through SAW excitation; protein electrophoresis shows there is less than 1% degradation of either protein molecule in this process. Finally, a solution of live yeast cells was infused into paper, which was subsequently dried for preservation then remoistened to extract the cells viaSAW atomization, yielding live cells at the completion of the process. The successful preservation and extraction of fluids, proteins and yeast cells significantly expands the usefulness of paper in microfluidics. [ABSTRACT FROM AUTHOR]

This article describes FLASH (Fast Lithographic Activation of Sheets), a rapid method for laboratory prototyping of microfluidic devices in paper. Paper-based microfluidic devices are emerging as a new technology for applications in diagnostics for the developing world, where low cost and simplicity are essential. FLASH is based on photolithography, but requires only a UV lamp and a hotplate; no clean-room or special facilities are required (FLASH patterning can even be performed in sunlight if a UV lamp and hotplate are unavailable). The method provides channels in paper with dimensions as small as 200 µm in width and 70 µm in height; the height is defined by the thickness of the paper. Photomasks for patterning paper-based microfluidic devices can be printed using an ink-jet printer or photocopier, or drawn by hand using a waterproof black pen. FLASH provides a straightforward method for prototyping paper-based microfluidic devices in regions where the technological support for conventional photolithography is not available. [ABSTRACT FROM AUTHOR]

In the present study, iodide-mediated 3,3',5,5'-tetramethylbenzidine (TMB)-H 2 O 2 reaction system was applied to a microfluidic paper-based analytical device (μPAD) for non-enzymatic colorimetric determination of H 2 O 2 . The proposed system is portable and incorporates a μPAD with a machine learning-based smartphone app. A smartphone app called " Hi-perox Sens " capable of image capture, cropping and processing was developed to make the system simple and user-friendly. Briefly, circular μPADs were designed and tested with varying concentrations of H 2 O 2 . Following the color change, the images of the μPADs were taken with four different smartphones under seven different illumination conditions. In order to make the system more robust and adaptive against illumination variation and camera optics, the images were first processed for feature extraction and then used to train machine learning classifiers. According to the results, TMB + KI showed the highest classification accuracy (97.8%) with inter-phone repeatability at t = 30 s under versatile illumination and maintained its accuracy for 10 minutes. In addition, the performance of the system was also comparable to two different commercially available H 2 O 2 kits in real samples.

Detection and characterization of DNA damage plays a critical role in genotoxicity testing, drug screening, and environmental health. We developed a fully integrated origami paper-based analytical device (oPAD) for measuring DNA damage. This simple device allows on-paper cell lysis, DNA extraction, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) reaction and signal readout with simple operation steps, enabling rapid (within 30 min) and high throughput assessment of multiple DNA damages induced by exogenous chemical agents.

Microfluidic paper-based analytical devices (μPADs) have been widely explored for point-of-care testing due to their simplicity, low cost, and portability. μPADs with multiple-step reactions usually require precise flow control, especially flow-delay. This paper reports the numerical, mathematical, and experimental studies of flow delay through wax valves surrounded by PDMS walls on paper microfluidics. The predried surfactant in the sample zone diffuses into the liquid sample which can therefore flow through the wax valves. The delay time is automatically regulated by the diffusion of the surfactant after sample loading. The numerical study suggested that both the elevated contact angle and the reduced porosity and pore size in the wax printed region could effectively prevent water but allow liquids with lower contact angles ( e.g. , surfactant solutions) to flow through. The PDMS walls fabricated using a low-cost liquid dispenser effectively prevented the leakage of surfactant solutions. By controlling the quantity, diffusion distance, and type of the surfactant predried on the chip, the system successfully achieved a delay time ranging from 1.6 to 20 minutes. A mathematical model involving the above parameters was developed based on Fick's second law to predict the delay time. Finally, the flow-delay systems were applied in sequential mixing and distance-based detection of either glucose or alcohol. Linear ranges of 1-100 mg dL -1 and 1-40 mg dL -1 were achieved for glucose and alcohol, respectively. The lower limit detection (LOD) of glucose and alcohol was 1 mg dL -1 . The LOD of glucose was only 1/11 of that detected using μPADs without flow control, indicating the advantage of controlling fluid flow. The systematic findings in this study provide critical guidelines for the development and applications of wax valves in automatic flow delay for point-of-care testing.

A novel miniaturized and sustainable platform exploiting two merged paper-based substrates has been applied for the programmable pre-concentration of analytes of interest and electrochemical detection of mercury traces in drinking water using printable sensor strips. This strategy represents a novel versatile possibility in merging humble materials maximizing their impacts on analytical and remediation challenges.

Leukocyte esterase (LE) is a useful marker that can be used in establishing a diagnosis of urinary tract infections (UTIs). The development of a UTI diagnostic method with quantitative determinations of biomarkers across all age groups is becoming more important. In this report, microfluidic resistance sensors based on silver ink (Ag ink) and silver ink mixed with ZnO nanoparticles (Ag-ZnO ink) were synthesized and coated on cellulose paper, namely LE-Ag-μPADs and LE-Ag-ZnO-μPADs, respectively, for the sensitive detection of LE. The microfluidic design increases the precision of data and further allows for quantitative determination and early detection of LE in human urine. The quantification of LE relies on the change in the resistance readout coating with Ag ink as well as Ag-ZnO ink in the detection zone. A mixture of 3-(N-tosyl-l-alaninyloxy)-5-phenylpyrrole (PE) and 1-diazo-2-naphthol-4-sulfonic acid (DAS) was deposited in the sample zone to selectively recognize LE, and the resulting nonconductive products, i.e., azo compounds, further reacted with the Ag ink and Ag-ZnO ink to increase resistance. The quantitative detectable LE concentrations between 2 to 32 (×5.2 U mL-1), i.e. ≈12 to 108 μg L-1, cover the commercial dipstick range of trace, +1 and +2. The minimum detectable concentration of LE in urine was 1 (×5.2 U mL-1). The lower concentrations of LE detectable by LE-Ag-μPADs (1-8 × 5.2 U mL-1) are below the value achieved with the ELISA LE kit. Urine samples from inpatients with indwelling urinary catheters were used, and the LE levels measured by the present device were highly correlated with those determined by a commercial urine analyser.

Negative ion mode paper spray mass spectrometry (PS-MS) suffers from intense background noise and unstable MS signal. For the first time, we reported fluorinated boron nitride nanosheet (h-FBN) assisted negative ion PS-MS for the detection of a series of molecules. We demonstrated that the introduction of h-FBN can greatly improve the detection sensitivity and signal stability in the negative ion mode.

Infectious diseases cause millions of deaths annually in the developing world. Recently, microfluidic paper-based analytical devices (μPADs) have been developed to diagnose such diseases, as these tests are low cost, biocompatible, and simple to fabricate. However, current μPADs are difficult to use in resource-limited areas due to their reliance on external instrumentation to measure and analyze the test results. In this work, we propose an electricity and external instrumentation-free μPAD sensor based on the colorimetric enzyme-linked immunosorbent assay (ELISA) for the diagnosis of infectious disease (3D-tPADs). Designed based on the principle of origami, the proposed μPAD enables the sequential steps of the colorimetric ELISA test to be completed in just ∼10 min. In addition, in order to obtain an accurate ELISA result without using any instrument, we have integrated an electricity-free "timer" within the μPAD that can be controlled by the buffer viscosity and fluid path volume to indicate the appropriate times for washing and color development steps, which can avoid false positive or false negative results caused by an extended or shortened amount of washing and development times. Due to the low background noise and high positive signal intensity of the μPAD, positive and negative detection results can be distinguished by just the naked eye. Furthermore, the ELISA result can be semi-quantified by comparing the results shown on the μPAD with a color chart diagram with a detection limit of HIV type 1(HIV-1) p24 antigen as low as 0.03 ng mL-1. These results demonstrate the proposed sensor can perform infectious disease diagnosis without external instrumentation or electricity, extending the application of the μPAD test for on-site detection and use in resource-limited settings.

Lab-on-paper, or microfluidic paper-based analytical devices (μPADs), use paper as a substrate material, and are patterned with a system of microchannels, reaction zones and sensing elements to perform analysis and detection. The sample transfer in such devices is performed by capillary action. As a result, external driving forces are not required, and hence the size and cost of the device are significantly reduced. Lab-on-paper devices have thus attracted significant attention for point-of-care medical diagnostic purposes in recent years, particularly in less-developed regions of the world lacking medical resources and infrastructures. This review discusses the major advances in lab-on-paper technology for blood analysis and diagnosis in the past five years. The review focuses particularly on the many clinical applications of lab-on-paper devices, including diabetes diagnosis, acute myocardial infarction (AMI) detection, kidney function diagnosis, liver function diagnosis, cholesterol and triglyceride (TG) analysis, sickle-cell disease (SCD) and phenylketonuria (PKU) analysis, virus analysis, C-reactive protein (CRP) analysis, blood ion analysis, cancer factor analysis, and drug analysis. The review commences by introducing the basic transmission principles, fabrication methods, structural characteristics, detection techniques, and sample pretreatment process of modern lab-on-paper devices. A comprehensive review of the most recent applications of lab-on-paper devices to the diagnosis of common human diseases using blood samples is then presented. The review concludes with a brief summary of the main challenges and opportunities facing the lab-on-paper technology field in the coming years.

The article focuses on controlled transport of reagents within paper devices using two dimensional (2D) paper networks. The geometry of the paper network and dissolvable barriers makes them able to transport multiple analytes and reagents. It states these paper-based lateral flow immunoassay strips analyse only a single analyte per strip but their utility can be increased using highly sensitive particle labels like quantum dots. It also offers information on an experiment and its result.

The major obstacle to developing nanozymes which are considered as promising alternatives to natural enzymes is their moderate performance, including poor affinity for substrates, low catalytic activity, and severe pH-dependence. To address these issues, herein, we synthesize ultrathin layered double hydroxide (LDH) nanosheets with a thickness of 1.4 nm and an average lateral size of 23 nm using a fast-precipitation method. Through the rational design of their compositions, it is found that NiMn LDHs exhibit the optimum peroxidase mimicking performance with excellent substrate affinity, high catalytic activity (a limit of detection (LOD) of 0.04 μM H 2 O 2 ) and robustness in a wide pH range (from 2.6 to 9.0), which is superior to that of natural horseradish peroxidase (HRP). The main active centers are identified as Mn sites because of their strong Lewis acidity and low redox potential. Furthermore, a series of disposable paper bioassays based on NiMn LDH nanozymes are designed and used for the highly sensitive detection of H 2 O 2 and ascorbic acid (AA).

This work reports the development of a rapid, simple and inexpensive colorimetric paper-based assay for the detection of the severe acute respiratory symptom coronavirus 2 (SARS-CoV-2) humanized antibody. The paper device was prepared with lamination for easy sample handling and coated with the recombinant SARS-CoV-2 nucleocapsid antigen. This assay employed a colorimetric reaction, which is followed by horseradish peroxidase (HRP) conjugated detecting antibody in the presence of the 3,3',5,5'-tetramethylbenzidine (TMB) substrate. The colorimetric readout was evaluated and quantified for specificity and sensitivity. The characterization of this assay includes determining the linear regression curve, the limit of detection (LOD), the repeatability, and testing complex biological samples. We found that the LOD of the assay was 9.00 ng μL-1 (0.112 IU mL-1). The relative standard deviation was approximately 10% for a sample number of n = 3. We believe that our proof-of-concept assay has the potential to be developed for clinical screening of the SARS-CoV-2 humanized antibody as a tool to confirm infected active cases or to confirm SARS-CoV-2 immune cases during the process of vaccine development.

Here, we present a new diacetoxy-functionalized UiO-66 metal-organic framework (MOF) for the trace level detection of hydrazine in water. The MOF material (1) was solvothermally prepared by the reaction between ZrOCl2·8H2O and 2,5-diacetoxy-1,4-benzenedicarboxylic acid (H2BDC-(OCOCH3)2). The desolvated material (1') showed a highly selective fluorescent turn-on signal towards hydrazine in water, which can be visualized by the naked eye under a UV lamp. Within 1 min of hydrazine addition, there was 14-fold fluorescence enhancement. The probe can detect hydrazine up to the nanomolar level (detection limit = 78.8 nM) in water. This detection limit is the lowest among MOF-based fluorescent probes for hydrazine. The material was also utilized for the sensing of hydrazine in paper strips and environmental water samples. Hydrazine-selective deprotection of ester groups anchored with the ligand is the principal reason behind the switch-on nature of sensing.

The SARS-CoV-2 virus is known as the causal agent for the current COVID-19 global pandemic. The majority of COVID-19 patients develop acute respiratory distress syndrome (ARDS), while some experience a cytokine storm effect, which is considered as one of the leading causes of patient mortality. Lipids are known to be involved in the various stages of the lifecycle of a virus functioning as receptors or co-receptors that controls viral propagation inside the host cell. Therefore, lipid-related metabolomics aims to provide insight into the immune response of the novel coronavirus. Our study has focused on determination of the potential metabolomic biomarkers utilizing a Teslin® Substrate in paper spray mass spectrometry (PS-MS) for the development of a rapid detection test within 60 seconds of analysis time. In this study, results were correlated with PCR tests to reflect that the systemic responses of the cells were affected by the COVID-19 virus.

This study describes a microfluidic paper-based analytical device (μPAD) for separating plasma from whole blood and measuring glucose concentration. A two-dimensional μPAD was fabricated by wax printing, using chromatographic paper and was functionalized by the incorporation of chitosan into the plasma separation zone. Red blood cells were isolated from whole blood in the plasma separation zone consisting of a chitosan polymer structure and a wax barrier. The separated plasma, with buffer, was injected into the test zone to cause a color change. The colorimetric results were digitized with a scanner, and the concentration was calculated using a calibration curve. This μPAD, which separated the plasma from whole blood without a separation membrane, will be useful for constructing point-of-care testing devices that detect analytes in small sample volumes.

Fibrinogen (FIB) plays a key role in blood coagulation and thrombosis and its concentration in blood can directly reflect health conditions, thus efficient detection of FIB would benefit the treatments of certain diseases such as liver and heart diseases. However, there is a lack of sensitive, simple, rapid and cheap FIB detection device currently, in lieu of expensive and sophisticated approaches in laboratories. Here, we propose a novel plasma separation and FIB detection platform based on a microfluidic paper-based analytical device (μPAD). It is the first time that dielectrophoretic (DEP) force is combined with capillary force on paper for plasma separation, and the separation efficiency of plasma reaches about 95%, ensuring reliable downstream FIB detection, for which we also propose a new method called the resistance-fibrinogen detection (RFD) method. It not only avoids the use of large-scale instruments for detection, but also possesses high precision and simplicity. The method is found to be reliable in FIB detection for various concentrations ranging from 127.0 to 508.0 mg dL -1 . Moreover, the results obtained from the proposed μPAD show an excellent agreement (R 2 = 0.9985) with those obtained from an automatic coagulation analyzer with natural human blood samples. Overall, the proposed platform provides a low-cost and reliable approach for FIB detection, especially for clinical use in resource-limited areas.

Globally, rapid development of antibiotic resistance amongst pathogens has led to limited treatment options and high indirect costs to health management. There is a need to avoid misuse of available antibiotics and to develop rapid, affordable and accessible diagnostic technologies to detect drug resistance even in resource limited settings. This study reports the development of instrument-free point-of-care devices for detection of antibiotic resistance for rapid diagnosis of drug resistance in the penicillin, cephalosporin and carbapenem groups of antibiotics. The simple paper-based devices for flow through assay determine the presence of resistant bacteria in a sample by a visible colour change within 30 minutes. At the center of this technology is the unique sensing nanomaterial comprising of core-shell nanoparticles layered with specific antibiotics. The core is comprised of chitosan nanoparticles of size ∼15 nm coated with the starch-iodine indicator to form a shell increasing the size to ∼47 nm. The test strip is coated with the nanoparticles, air-dried and overlayed with the required antibiotic. In the presence of penicillin, cephalosporin and carbapenem resistant bacteria, the core-shell nanoparticles undergo a visible colour change from blue to white. The core-shell nanoparticles were deposited on paper to form a point-of-care device. Devices were developed to screen for three main classes of antibiotics namely penicillins, cephalosporins and carbapenems. The devices were validated using standard resistant and susceptible ATCC strains in three different sample types, pure colony, broth culture and saline suspensions. The change of colour from blue to white was considered a positive test. The time of detection was found to be 30 min, while the limit of detection was 10 5 cfu ml -1 . The device exhibited 100% sensitivity and specificity with known resistant and susceptible cultures not only from pure colonies but also from direct samples of spiked saline suspensions with graded confounding factors of albumin, glucose, and urea. The inter-device reproducibility and storage stability of the devices was established. The developed point-of-care devices have potential as screening devices for antimicrobial resistance.

We report for the first time a highly sensitive and rapid quantitative method for the detection of Salmonella Typhimurium (S. Typhimurium) using a conductive immunosensor on a paper-based device (PAD). S. Typhimurium monoclonal antibodies (MA) were first immobilized on a paper-based device and then captured by S. Typhimurium. After an immunoreaction on the device, the polyclonal antibody-colloidal gold conjugate (PA-AuNPs) was dropped to bind with S. Typhimurium. After a complete sandwich reaction, a dark red color appeared on the paper-based device, which can be observed by the naked eye for a rapid screening test. The electrical conductivity of PA-AuNPs between the screen-printed electrodes on the paper-based device was also measured for an accurate quantitative analysis. The electrical conductivity correlated well with the concentration of S. Typhimurium, which was controlled by the amount of S. Typhimurium attached to the polyclonal antibody-colloidal gold conjugate. The device showed a linear correlation for the concentration of the S. Typhimurium in the range of 10-10 8 CFU mL -1 in a logarithmic plot, with an R 2 value of 0.9882 and a limit of detection (LOD) as low as 10 CFU mL -1 . This simple, highly sensitive, and rapid method for the S. Typhimurium detection was successfully performed within 30 min, and it can be developed into small portable measuring devices in order to facilitate preliminary screening tests.

An increasing number of patients are living with Alzheimer's disease (AD); thus, the need for a method to detect AD early and sensitively has become urgent, and the demand for an intelligent analytical platform is growing year by year. Abnormal levels of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are known to be indicative of AD. In this work, a novel conjugated polythiophene (CP) compound was successfully combined with CdTe quantum dots (QDs) to improve their selectivity and sensitivity. The QDs successfully enabled the detection of low concentrations of AChE by turning on the fluorescence of the CdTe/CP via the interaction between CP and thiocholine produced by ATCh hydrolysis and aggregation induced emission enhancement (AIEE). Under optimal conditions, we reached a low detection limit of 0.14 U L -1 , which is 7.9 times lower than that of pristine QDs. More importantly, an efficient, inexpensive, and disposable paper-based platform, which allows the efficient visual detection of AChE activity via the color variation of CdTe/CP, was designed. Moreover, the accuracy of the method was demonstrated by conducting a recovery test in human serum, in which the recoveries reached 107% and 110%, proving that CdTe/CP has considerable potential to be used for analyzing real biological samples. The advantages of this method are its simplicity, fast detection capability, affordability, and the fact that it can be used for on-site detection of AChE activity. Furthermore, it has certain guiding significance for detecting AD.

In this paper, a paper-supported analytical device based on a sandwich immunoreaction and luminescence resonance energy transfer (LRET) was reported for the visual and quantitative determination of a cancer biomarker, in which upconversion nanoparticles (UCNPs) were located on the surface of the paper as energy donors and gold nanoparticles (AuNPs) were used as energy acceptors. Upon the recognition of the cancer biomarker by two rationally selected antibodies, the upconversion luminescence was quenched by the AuNPs in a biomarker concentration-dependent manner. As a model target, CEA was detected using this immunosensor, and a linear relationship within 0.5-30 ng mL-1 was obtained in buffer solution, with a detection limit of 0.21 ng mL-1. The immunosensor was also applicable in 20-fold diluted human serum with a linear range of 0.5-30 ng mL-1 and a detection limit of 0.36 ng mL-1. This technique also realized the qualitative judgment of the critical concentration of CEA in serum samples by the naked eye. This approach displays great application potential for point-of-care testing in clinical applications, as well as the potentiality to be extended to other kinds of disease-related biomolecules.

The first discrete bimetallic europium(iii)/copper(ii) complex for the fast, sensitive and selective luminescent detection of both aqueous and gaseous hydrogen sulfide has been developed. The chemosensor displayed an impressive response time of 30 seconds and a low theoretical limit of detection (100 ppb) for gaseous hydrogen sulfide.

In the present article we report a new hybrid microfluidic device (hyFlow) comprising a disposable paper electrode and a three-dimensional (3D) printed plastic chip for the electrochemical detection of a magnetic bead-silver nanoparticle (MB-AgNP) bioconjugate. This hybrid device evolved due to the difficulty of incorporating micron-scale MBs into paper-only fluidic devices. Specifically, paper fluidic devices can entrap MB-containing conjugates within their cellulose or nitrocellulose fiber matrix. The hyFlow system was designed to minimize such issues and transport MB conjugates more efficiently to the electrochemical detection zone of the device. The hyFlow system retains the benefit of fluid transport by pressure-driven flow, however, no pump is required for its operation. The hyFlow device is capable of detecting either pre-formed MB-AgNP conjugates or conjugates formed in situ. The detection limit of AgNPs using this device is 12 pM, which represents just 22 AgNPs per MB.

Recently, portable sensing devices with point of care testing (POCT) capability have attracted great attention due to their inherent affordability and accessibility in low resource areas. Paper sensors possess excellent potential as POCT platforms because of low cost, ease of operation, disposability and high-volume manufacturing. Paper sensors that incorporate functional nucleic acids (FNAs) as recognition elements are particularly attractive given that FNAs can be isolated from random-sequence nucleic acid pools to recognize, or respond to, virtually any target of interest. In this review, the advantages of FNAs, particularly DNA aptamers and DNAzymes, as recognition elements for the design of paper sensors are first discussed. This is followed by reviewing three specific types of FNA based paper sensors: dot blots, lateral flow assays, and microfluidic paper-based analytical devices. Furthermore, advances in the signal reporting methods used by FNA based paper sensors are summarized. Finally, limitations of current FNA based paper sensors are discussed along with considerations of future research directions.

Atherosclerosis (AS) is the most common factor causing many cardiovascular and cerebrovascular diseases and has received considerable attention. The occurrence mechanism of AS is uncertain because it is a choronically pathological process that is influenced by multi-aspects, among which cytokines play the key roles in regulating the processes of the immune system. For example, two key cytokines, namely, IL-10 and MCP-1 (chemokine), which are involved in AS progression with varied levels, can be used for AS status monitoring and early diagnosis of AS-associated diseases. Hence, a new paper-based, surface-enhanced Raman spectroscopy (SERS) sensing platform was established for the detection of these two key cytokines. By combining a nanoporous networking membrane as the substrate and SERS nanotags as the probe for signal reading, together with a sandwich design, sensitive and specific identification and quantification of cytokine targets in human serum were achieved with excellent sensing characteristics. The lowest detectable concentration was determined to be 0.1 pg mL-1 for both IL-10 and MCP-1 in human serum. The assay also exhibits high specificity towards target cytokine detection, with low-nonspecific binding and acceptable cross-reactivity in the presence of other structurally similar targets. Finally, the practicability was validated by performing duplex detection in human serum, which further demonstrates the high specificity of the assay for the detection of target cytokines. Taken together, these promising results illustrate that this developed sensing assay is a candidate for clinical multi-target analysis in real environments.

The low solubility of gas molecules in aqueous solutions has limited the power density output of enzymatic biofuel cells. Herein, a single-liquid miniature glucose-O2 fuel cell was constructed by using gas diffusion electrodes, which were prepared by immobilizing glucose oxidase (GOx) or laccase (Lac) modified on a porous structured carbon paper (CP). Due to the fast and direct O2 diffusion from air to the active sites of the immobilized enzyme through the pores of the CP anode/cathode with controlled wettability, the maximum power output densities dramatically increased to 9.64 μW cm-2 at 0.43 V and 53.0 μW cm-2 at 0.45 V for the cell in 5 mM glucose and after exposing the cell to air or O2 atmosphere, respectively. Interestingly, the resulting single-liquid cell could harvest power from human serum operating at a maximum power density of 49.0 μW cm-2 at 0.2 V. The biofuel cell fabricated by the gas diffusion electrodes displayed advantages such as high output power density, low cost and high 'on-chip' integrability and miniaturization, which suggest its great potential for implantable self-powered sensors and for many future applications.