Development of a C-reactive protein quantification method based on flow rate measurement of an ink solution pushed out by oxygen gas generated by catalase reaction.

A novel determination method for protein biomarkers based on on-chip flow rate measurement was developed using a microchip with organic photodiodes (OPDs). This quantitative method is based on the flow rate measurement of an ink solution pushed out by oxygen gas generated through catalase reaction. The amount of oxygen gas generated in the sample reservoir is dependent on the concentration of the analyte; therefore, the flow rate of the ink solution is also dependent on the concentration of the analyte. The concentration of the analyte can thus be estimated by measurement of the ink solution flow rate. The ink solution flow rate was estimated by measuring the migration time of the ink solution between two points using two OPDs placed below the microchannel. The principle of this method was demonstrated by the measurement of catalase using the microchip. In addition, the developed method was applied to the determination of C-reactive protein (CRP), a biomarker of inflammation, based on a catalase-linked immunosorbent assay (C-LISA). The limit of detection for CRP was 0.20 µg/mL. The method was also applied to the determination of CRP in human serum, and the quantitative values obtained by this method were in excellent agreement with those obtained by the conventional enzyme-linked immunosorbent assay (ELISA) method. The developed method does not require a photodetector with high sensitivity and is thus capable of downsizing; therefore, this will be useful for on-site analyses such as point-of-care testing and field measurements.

Rapid measurement of bacterial contamination in water: A catalase responsive-electrochemical sensor.

The present study describes the development of a potentiometric sensor for microbial monitoring in water based on catalase activity. The sensor comprises a MnO2-modified electrode that responds linearly to hydrogen peroxide (H2O2) from 0.16 M to 3.26 M. The electrode potential drops when the H2O2 solution is spiked with catalase or catalase-producing microorganisms that decompose H2O2. The sensor is responsive to different bacteria and their catalase activities. The electrochemical sensor exhibits a lower limit of detection (LOD) for Escherichia coli at 11 CFU/ml, Citrobacter youngae at 12 CFU/ml, and Pseudomonas aeruginosa at 23 CFU/ml. The sensor shows high sensitivity at 3.49, 3.02, and 4.24 mV/cm2dec for E. coli, C. youngae, and P. aeruginosa, respectively. The abiotic sensing electrode can be used multiple times without changing the response potential (up to 100 readings) with a shelf-life of over six months. The response time is a few seconds, with a total test time of 5 min. Additionally, the sensor effectively tested actual samples (drinking and grey water), which makes it a quick and reliable sensing tool. Therefore, the study offers a promising water monitoring tool with high sensitivity, stability, good detection limit, and minimum interference from other water contaminants.