A Paper Sensor Printed with Multifunctional Bio/Nano Materials.

We report a paper-based aptasensor platform that uses two reaction zones and a connecting bridge along with printed multifunctional bio/nano materials to achieve molecular recognition and signal amplification. Upon addition of analyte to the first zone, a fluorescently labelled DNA or RNA aptamer is desorbed from printed graphene oxide, rapidly producing an initial fluorescence signal. The released aptamer then flows to the second zone where it reacts with printed reagents to initiate rolling circle amplification, generating DNA amplicons containing a peroxidase-mimicking DNAzyme, which produces a colorimetric readout that can be read in an equipment-free manner or with a smartphone. The sensor was demonstrated using an RNA aptamer for adenosine triphosphate (a bacterial marker) and a DNA aptamer for glutamate dehydrogenase (Clostridium difficile marker) with excellent sensitivity and specificity. These targets could be detected in spiked serum or feacal samples, demonstrating the potential for testing clinical samples.

Paper-based assay for red blood cell antigen typing by the indirect antiglobulin test.

A rapid and simple paper-based elution assay for red blood cell antigen typing by the indirect antiglobulin test (IAT) was established. This allows to type blood using IgG antibodies for the important blood groups in which IgM antibodies do not exist. Red blood cells incubated with IgG anti-D were washed with saline and spotted onto the paper assay pre-treated with anti-IgG. The blood spot was eluted with an elution buffer solution in a chromatography tank. Positive samples were identified by the agglutinated and fixed red blood cells on the original spotting area, while red blood cells from negative samples completely eluted away from the spot of origin. Optimum concentrations for both anti-IgG and anti-D were identified to eliminate the washing step after the incubation phase. Based on the no-washing procedure, the critical variables were investigated to establish the optimal conditions for the paper-based assay. Two hundred ten donor blood samples were tested in optimal conditions for the paper test with anti-D and anti-Kell. Positive and negative samples were clearly distinguished. This assay opens up new applications of the IAT on paper including antibody detection and blood donor-recipient crossmatching and extends its uses into non-blood typing applications with IgG antibody-based diagnostics. Graphical abstract A rapid and simple paper-based assay for red blood cell antigen typing by the indirect antiglobulin test.

The detection of blood group phenotypes using paper diagnostics.

BACKGROUND AND OBJECTIVES: Paper biodiagnostics for blood typing are novel, cheap, fast and easy to use. Agglutinated red blood cells cannot travel through the porous structure of paper, indicating a positive antibody-antigen interaction has occurred. Conversely, non-agglutinated blood can disperse and wick through the paper structure with the ease to indicate a negative result. This principle has been demonstrated to detect blood group phenotypes: ABO and RhD. However, typing for red blood cell antigens such as Rh, Kell, Duffy and Kidd has not yet been explored on paper. MATERIALS AND METHODS: Two paper testing methods - an elution and a direct flow-through method - were investigated to detect red blood cell antigens excluding the ABO system and RhD. Antigens explored include the following: C, c, E, e, K, k, Fy(a), Fy(b), Jk(a), Jk(b), M, N, S and s, P1, Le(a) and Le(b). The variables tested include the following: reaction time and reagent concentration. The importance of antibody type/structure for successful agglutination on paper was confirmed. RESULTS: Some blood group phenotypes showed less agglutination due to weaker antibody-antigen interactions. Most blood groups with antibodies available as IgM, such as C, c, E, e, K and k, and Jk(a) and Jk(b), and P1, were successful using both methods. However, other blood groups, especially those with antibodies only available as polyclonal antibodies, were unsuccessful and require further scrutiny. CONCLUSION: Paper can be used as an alternative blood grouping diagnostic tool for selected blood group phenotypes.

Functional Comparison of Bioactive Cellulose Materials Incorporating Engineered Binding Proteins.

Whatman No. 1 chromatography paper is widely used as a substrate for cellulose-based immunoassays. The immobilized proteins are used to capture target biomarkers for detection. However, alternative paper substrates may facilitate mass production of immunoassays as diagnostic tests. Here, we assessed the physical characteristics and protein immobilization capabilities of different commercial papers. Some substrates fulfilled our design criteria, including adequate flow rate and sufficient protein immobilization for efficient target capture. This study demonstrates that a variety of paper substrates can be bioactivated and used to capture target biomarkers, enabling development of affordable diagnostic tests from a range of starting materials.

Combined optical and electronic paper-nose for detection of volatile gases.

In this work, a paper-based optoelectronic sensor (paper-nose) is presented for sensing volatile gases in air. The proposed optoelectronic sensor is a combination of both colorimetric (optical) and chemiresistive (electronic) sensor arrays in order to improve the selectivity of the paper-nose in the complex air background. The optical sensors are based on chemoresponsive dyes, namely Reichardt's dye (2,6-diphenyl-4-(2,4,6-triphenyl-1-pyridinio)phenolate), bromocresol purple, methyl red, bromothymol blue, brilliant yellow and manganese tetraphenylporphyrin (Mn-TPP). The chemiresistive sensors are based on nanomaterials, such as carbon nanotubes (CNT), PEDOT:PSS, graphite, and an ionic liquid, 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide (EMI TFSI). Sensor is fabricated through direct handwriting of sensing materials using a pen on paper without the need of expensive cleanroom facilities. The optoelectronic sensor is tested in ambient air with different volatile gases such as methanol, ammonia, toluene, acetone and ethanol and their mixtures of varying concentrations. The detected electrical and optical responses together form a unique signature for each volatile gas and its mixture. Support-vector machine (SVM) is applied for target classification and detection. From the SVM result, it is found that better discriminative power is achieved by combining optical and electrical responses.

Developing Paper Based Diagnostic Technique to Detect Uric Acid in Urine.

Urinary or serum uric acid concentration is an indicator of chronic kidney condition. An increase in uric acid concentration may indicate renal dysfunction. Reliable instantaneous detection of uric acid without requiring sophisticated laboratory and analytical instrumentation, such as: chromatographic and spectrophotometric methods, would be invaluable for patients with renal complication. This paper reports the early development of a simple, low-cost, instantaneous and user-friendly paper based diagnostic device (PAD) for the qualitative and quantitative detection of uric acid in urine. A colorimetric detection technique was developed based on the intensity of Prussian blue color formation on paper in presence of uric acid; the reaction rate of corresponding chemical reactions on paper surface was also studied. Based on the colorimetric signal produced on paper surface, a calibration curve was developed to detect unknown concentration of uric acid in urine. The effect of temperature on formation of color signal on paper surface was also analyzed. In this study, estimation of urinary uric acid using MATLAB coding on a windows platform was demonstrated as the use of software application and digital diagnostics. This paper-based technique is faster and less expensive compared to traditional detection techniques. The paper-based diagnostic can be integrated with a camera of smart phone, tablet computer or laptop and an image processing application (using windows/android/IOS platform) as a part of digital diagnostics. Therefore, with proper calibration, the paper-based technique can be compatible and economical to the sophisticated detection techniques used to detect urinary uric acid.

Sensitive Protein Detection and Quantification in Paper-Based Microfluidics for the Point of Care.

The design of appropriate diagnostic assays for the point of care requires development of suitable biosensors, detection methods, and diagnostic platforms for sensitive, quantitative detection of biological analytes. Protein targets in particular are especially challenging to detect quantitatively and sensitively due to the lack of amplification strategies akin to nucleic acid amplification. However, recent advances in transducer and biosensor design, new detection labels, and paper-based microfluidics may realize the goal of sensitive, fast, portable, and low-cost protein detection. In this review, we discuss the biochemistry, optics, and engineering advances that may be leveraged to design such a sensitive protein diagnostic assay. The binding kinetics, mechanisms of binding in porous networks, and potential transducers are explained in detail. We discuss the relative merits of various optical detection strategies, potential detection labels, optical readout approaches, and image-processing techniques that are amenable to point-of-care use. To conclude, we present a systematic analysis of potential approaches to enhance the sensitivity of paper-based assays. The assay development framework presented here provides bioassay developers a strategy to methodically enhance the sensitivity and point-of-care suitability of protein diagnostics.

Qualitative and quantitative detection of T7 bacteriophages using paper based sandwich ELISA.

Viruses cause many infectious diseases and consequently epidemic health threats. Paper based diagnostics and filters can offer attractive options for detecting and deactivating pathogens. However, due to their infectious characteristics, virus detection using paper diagnostics is more challenging compared to the detection of bacteria, enzymes, DNA or antigens. The major objective of this study was to prepare reliable, degradable and low cost paper diagnostics to detect viruses, without using sophisticated optical or microfluidic analytical instruments. T7 bacteriophage was used as a model virus. A paper based sandwich ELISA technique was developed to detect and quantify the T7 phages in solution. The paper based sandwich ELISA detected T7 phage concentrations as low as 100 pfu/mL to as high as 10(9) pfu/mL. The compatibility of paper based sandwich ELISA with the conventional titre count was tested using T7 phage solutions of unknown concentrations. The paper based sandwich ELISA technique is faster and economical compared to the traditional detection techniques. Therefore, with proper calibration and right reagents, and by following the biosafety regulations, the paper based technique can be said to be compatible and economical to the sophisticated laboratory diagnostic techniques applied to detect pathogenic viruses and other microorganisms.