Thyroid stimulating hormone (TSH) plays a critical role in regulating physiological symptoms in human, thus it is important to detect TSH with high sensitivity and in human serum sample. In this study, we proposed a strategy to detect TSH antigen using a membrane-based microwave-mediated analytical device developed for highly responsive electrochemical immunoassay with sandwich ELISA format. Experimental assay was done with different TSH antigen concentrations. It was found that the immunoassay was able to detect TSH antigen as low as 0.2 mIU/L. The total detection time from preparation of samples to measurement was within 60 min. The specificity of this immunoassay was proven that against unspecific proteins as the almost low signal was obtained for all tested solutions. Finally, exploratory experiments using standard human serum samples with TSH antigens were carried out prior to a successful trial using clinical human serum samples. Our results indicate that this membrane-based microwave-mediated analytical electrochemical immunoassay device could be used as a new platform for low cost, sensitive, specific, swift, and point-of-care diagnosis of TSH. Moreover, this advanced system could define a powerful point-of-care tool in the next generation of technology in detecting and signaling biomarkers at early stages by using electrochemical immunosensor.

Here, we report a simple and effective method for capturing and displacement of gram-negative bacteria using aptamer-modified microbeads and acoustophoresis. As acoustophoresis allows for simultaneous washing and size-dependent separation in continuous flow mode, we efficiently obtained gram-negative bacteria that showed high affinity without any additional washing steps. The proposed device has a simple and efficient channel design, utilizing a long, square-shaped microchannel that shows excellent separation performance in terms of the purity, recovery, and concentration factor. Microbeads (10 µm) coated with the GN6 aptamer can specifically bind gram-negative bacteria. After incubation of bacteria culture sample with aptamer affinity bead, gram-negative bacteria-bound microbeads, and other unbound/contaminants can be separated by size with high purity and recovery. The device demonstrated excellent separation performance, with high recovery (up to 98%), high purity (up to 99%), and a high-volume rate (500 µL/min). The acoustophoretic separation performances were conducted using 5 Gram-negative bacteria and 5 Gram-positive bacteria. Thanks to GN6 aptamer’s binding affinity, aptamer affinity bead also showed binding affinity to multiple strains of gram-negative bacteria, but not to gram-positive bacteria. GN6 coated bead can capture Gram-negative bacteria but not Gram-positive bacteria. This study may present a different perspective in the field of early diagnosis in bacterial infectious diseases. In addition to detecting living bacteria or bacteria-derived biomarkers, this protocol can be extended to monitoring the contamination of water resources and may aid quick responses to bioterrorism and pathogenic bacterial infections.

A repeated usable electrode sensor for the measurement of glucose in urine has been developed. For improvement of the selectivity in measuring glucose in urine, we fabricated the Pt electrode coated with multilayered composite of glucose oxidase (GOX) and ascorbate oxidase (AOX). This simple layer structure could deliver more electrons the electrode from glucose oxidation without interferent from ascorbic acids to by firstly oxidizing the ascorbic acids present in urine. Results showed that our newly developed Pt-electrode is capable of quantifying the concentrations of glucose in urine min 0 to max 2,000mg/dL as well as maintaining its detection stability during two hundred times repeated measurements. This repeatable use of the sensor is attributable to the solidified structure of AOX and GOX layer.

The Enzyme Linked Immunosorbent Assay (ELISA) technique has been widely used for the identification and quantification of biochemical markers. The typical ELISA requires a number of washing steps to eliminate the unbound proteins which sometimes cause the desorption of protein due to their weak bonding between protein and well plate. In this study, we have developed a meshed type of plastic membrane in order to increase the reliable binding efficiency between proteins and the membrane surface, and to provide easy steps of washing. The use of our developed solid membrane has significantly increased the binding capacity of the biomolecules because this membrane ELISA (mELISA) provides 3D binding surfaces which increases the surface area when compared to the conventional 2D surface well plate. The columns were pretreated to form a self-assembled layer (SAM) on the surface for the stable conjugation of a target antibody. The SAM-coated membranes could be stored for one month without any further deterioration of stability. The measured optical density (O.D.) shows a 1.2-fold increase in IgG antigen (25 μg/mL) from the plastic membrane as compared with the conventional ELISA method. The concentrations of thyroid stimulating hormone were also monitored using the mELISA method and it shows good linearity against the concentrations.

Here, we report a simple and effective method for separation of gram-negative bacteria using aptamer-modified microbeads and acoustophoresis. As acoustophoresis allows for simultaneous washing and size-dependent separation in continuous flow mode, we efficiently obtained gram-negative bacteria that showed high affinity without any additional washing steps. The proposed device has a simple and efficient channel design, utilizing a long, square-shaped microchannel that shows excellent separation performance in terms of the purity, recovery, and concentration factor. Microbeads (10 µm) coated with the GN6 aptamer can specifically bind gram-negative bacteria. Using acoustophoresis, gram-negative bacteria-bound microbeads and other unbound/contaminants can be separated by size with high purity and recovery. The device demonstrated excellent separation performance, with high recovery (up to 98%), high purity (up to 99%), and a high volume rate (500 µL/min), and a concentration factor of up to 20×. The acoustophoresis microfluidic device also showed binding affinity to multiple strains of gram-negative bacteria, but not to gram-positive bacteria. This study presents a new paradigm for early diagnosis of bacterial infectious diseases. In addition to detecting living bacteria or bacteria-derived biomarkers, this protocol can be extended to monitoring the contamination of water resources, and may aid quick responses to bioterrorism and pathogenic bacterial infections.