In this paper, we represent a label-free biosensor based on immobilization of serum antibodies for rapid detection of viral antigens. Human serum containing specific antibodies against Japanese encephalitis virus (JEV) was immobilized on a silanized surface of an interdigitated sensor via protein A/glutaraldehyde for electrical detection of JEV antigens. The effective immobilization of serum antibodies on the sensor surface was verified by Fourier transform infrared spectrometry and fluorescence microscopy. The signal of the biosensor obtained by the differential voltage converted from the change into non-Faradic impedance resulting from the specific binding of JEV antigens on the surface of the sensor. The detection analyzed indicates that the detection range of this biosensor is 1-10 μg/ml JEV antigens, with a detection limit of 0.75 μg/ml and that stable signals are measured in about 20 min. This study presents a useful biosensor with a high selectivity for rapid and simple detection of JEV antigens, and it also proposes the biosensor as a future diagnostic tool for rapid and direct detection of viral antigens in clinical samples for preliminary pathogenic screenings in the case of possible outbreaks.
Pathogen separation is of great significance for precise detection and prevention of disease outbreaks. For the first time, protein A conjugated with chitosan-coated iron oxide nanoparticles was prepared for pathogen separation at low concentrations from liquid samples. Vibrio cholerae O1 (VO1) bacteria were used for testing the effectiveness of this conjugate. Transmission electron microscopy (TEM) was used to confirm the presence of captured VO1. The results showed that, after binding with a specific antibody, the conjugate allows separation of VO1 bacteria from water samples at a concentration as low as 10 cfu mL(-1). Moreover, the conjugate can be used in parallel with conventional or modern diagnostic tests for quick and accurate detection of pathogens.
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