Rapid Microfluidic Immuno-Biosensor Detection System for the Point-of-Care Determination of High-Sensitivity Urinary C-Reactive Protein.

A microfluidic immuno-biosensor detection system consisting of a microfluidic spectrum chip and a micro-spectrometer detection device is presented for the rapid point-of-care (POC) detection and quantification of high-sensitivity C-reactive protein (hs-CRP) in urine. The detection process utilizes a highly specific enzyme-linked immunosorbent assay (ELISA) method, in which capture antibodies and detection antibodies are pre-deposited on the substrate of the microchip and used to form an immune complex with the target antigen. Horseradish peroxidase (HRP) is added as a marker enzyme, followed by a colorimetric reaction using 3,3',5,5'-tetramethylbenzidine (TMB). The absorbance values (a.u.) of the colorimetric reaction compounds are measured using a micro-spectrometer device and used to measure the corresponding hs-CRP concentration according to the pre-established calibration curve. It is shown that the hs-CRP concentration can be determined within 50 min. In addition, the system achieves recovery rates of 93.8-106.2% in blind water samples and 94.5-104.6% in artificial urine. The results showed that the CRP detection results of 41 urine samples from patients with chronic kidney disease (CKD) were highly consistent with the conventional homogeneous particle-enhanced turbidimetric immunoassay (PETIA) method's detection results (R2 = 0.9910). The experimental results showed its applicability in the detection of CRP in both urine and serum. Overall, the results indicate that the current microfluidic ELISA detection system provides an accurate and reliable method for monitoring the hs-CRP concentration in point-of-care applications.

Microfluidic Sliding Paper-Based Device for Point-of-Care Determination of Albumin-to-Creatine Ratio in Human Urine.

A novel assay platform consisting of a microfluidic sliding double-track paper-based chip and a hand-held Raspberry Pi detection system is proposed for determining the albumin-to-creatine ratio (ACR) in human urine. It is a clinically important parameter and can be used for the early detection of related diseases, such as renal insufficiency. In the proposed method, the sliding layer of the microchip is applied and the sample diffuses through two parallel filtration channels to the reaction/detection areas of the microchip to complete the detection reaction, which is a simple method well suited for self-diagnosis of ACR index in human urine. The RGB (red, green, and blue) value intensity signals of the reaction complexes in these two reaction zones are analyzed by a Raspberry Pi computer to derive the ACR value (ALB and CRE concentrations). It is shown that the G + B value intensity signal is linearly related to the ALB and CRE concentrations with the correlation coefficients of R2 = 0.9919 and R2 = 0.9923, respectively. It is additionally shown that the ALB and CRE concentration results determined using the proposed method for 23 urine samples were collected from real suffering chronic kidney disease (CKD) patients are in fine agreement with those acquired operating a traditional high-reliability macroscale method. Overall, for point-of-care (POC) CKD diagnosis and monitoring in clinical applications, the results prove that the proposed method offers a convenient, real time, reliable, and low-spending solution for POC CKD diagnosis.

Microfluidic aptasensor POC device for determination of whole blood potassium.

An integrated microfluidic Au nanoparticle (AuNP) aptasensor device is proposed for monitoring the concentration of potassium (K+) ions in the bloodstream of patients with chronic kidney disease (CKD). In the proposed detection device, the AuNPs in the AuNP/aptamer complex are displaced by the serum K+ ions and react with NaCl to produce a color change in the detection region from which the K+ ion concentration is then inversely derived. The microfluidic device comprises two main components, namely an AuNP aptasensor PMMA (Poly(methyl methacrylate))/paper-microchip and a colorimetric analysis system for the quantitative detection of K+ ion concentration in whole blood. The functions of PMMA/paper microchips include reagent storage, K+ ion/aptamer reaction, and separation of serum from whole blood samples (blood filter). Experimental results show that the microfluidic device provides a linear response over the K+ ion concentration in range of 0.05-9 mM in artificial serum and has a detection limit (LOD) of 0.01 mM. Moreover, the detection results obtained for the 137 whole blood and 287 serum samples of CKD patients are very consistent (R2 = 0.968 and R2 = 0.980) with the measurement results obtained using an ion-selective electrodes (ISE) method. Results confirm that the current microfluidic aptasensor device provides a highly-sensitive and convenient method for performing the point-of-care (POC) monitoring of the whole blood K+ ion concentration.

Microfluidic colorimetric detection platform with sliding hybrid PMMA/paper microchip for human urine and blood sample analysis.

A microfluidic colorimetric detection (MCD) platform consisting of a sliding hybrid PMMA/paper microchip and a smart analysis system is proposed for the convenient, low-cost and rapid analysis of human urine and whole blood samples. The sliding PMMA/paper microchip comprises a PMMA microfluidic chip for sample injection and transportation, a paper strip for sample filtration (urine) or separation (blood), and a sealed paper-chip detection zone for sample reaction and detection. In the proposed device, the paper-chip is coated with bicinchoninic acid (BCA) and biuret reagent and is then assembled into the PMMA microchip and packaged in aluminum housing. In the detection process, the PMMA/paper microchip is slid partially out of the housing, and 2 µL of sample (urine or whole blood) is dripped onto the sample injection zone. The chip is then slid back into the housing and the sample is filtered/separated by the paper strip and transferred under the effects of capillary action to the sealed paper-chip detection zone. The housing is inserted into the color analysis system and heated at 45 °C for 5 min to produce a purple-colored reaction complex. The complex is imaged using a CCD camera and the RGB color intensity of the image is then analyzed using a smartphone to determine the total protein (TP) concentration of the sample. The effectiveness of the proposed method is demonstrated using TP control samples with known concentrations in the range of 0.03-5.0 g/dL. The detection results obtained for 50 human urine samples obtained from random volunteers are shown to be consistent with those obtained from a conventional hospital analysis system (R2 = 0.992). Moreover, the detection results obtained for the albumin (ALB) and creatine (CRE) concentrations of 50 whole blood samples are also shown to be in good agreement with the results obtained from the hospital analysis system (R2 = 0.982 and 0.988, respectively).

Microfluidic colorimetric analysis system for sodium benzoate detection in foods.

Sodium benzoate (SBA) is a widely-used additive for preventing food spoilage and deterioration and extending the shelf life. However, the concentration of SBA must be controlled under safe regulations to avoid damaging human health. Accordingly, this study proposes a microfluidic colorimetric analysis (MCA) system composing of a wax-printed paper-microchip and a self-made smart analysis equipment for the concentration detection of SBA in common foods and beverages. In the presented method, the distilled SBA sample is mixed with NaOH to obtain a nitro compound and the compound is then dripped onto the reaction area of the paper-microchip, which is embedded with two layers of reagents (namely acetophenone and acetone). The paper-microchip is heated at 120 °C for 20 min to cause a colorimetric reaction and the reaction image is then obtained through a CMOS (complementary metal oxide semiconductor) device and transmitted to a cell-phone over a WiFi connection. Finally, use the self-developed RGB analysis software installed on the cell-phone to obtain the SBA concentration. A calibration curve is constructed using SBA samples with known concentrations ranging from 50 ppm (0.35 mM) to 5000 ppm (35 mM). It is shown that the R + G + B value (Y) of the reaction image and SBA concentration (X) are related via Y = -0.034 X +737.40, with a determination coefficient of R2 = 0.9970. By measuring the SBA concentration of 15 commercially available food and beverage products, the actual feasibility of the current MCA system can be demonstrated. The results show that the difference from the measurement results obtained using the macroscale HPLC method does not exceed 6.0%. Overall, the current system provides a reliable and low-cost technique for quantifying the SBA concentration in food and drink products.