Paper-based electrochemical immunosensor for label-free detection of multiple avian influenza virus antigens using flexible screen-printed carbon nanotube-polydimethylsiloxane electrodes

Abstract

Many studies have been conducted on measuring avian influenza viruses and their hemagglutinin (HA) antigens via electrochemical principles; most of these studies have used gold electrodes on ceramic, glass, or silicon substrates, and/or labeling for signal enhancement. Herein, we present a paper-based immunosensor for label-free measurement of multiple avian influenza virus (H5N1, H7N9, and H9N2) antigens using flexible screen-printed carbon nanotube-polydimethylsiloxane electrodes. These flexible electrodes on a paper substrate can complement the physical weakness of the paper-based sensors when wetted, without affecting flexibility. The relative standard deviation of the peak currents was 1.88% when the electrodes were repeatedly bent and unfolded twenty times with deionized water provided each cycle, showing the stability of the electrodes. For the detection of HA antigens, approximately 10-μl samples (concentration: 100 pg/ml–100 ng/ml) were needed to form the antigen–antibody complexes during 20–30 min incubation, and the immune responses were measured via differential pulse voltammetry. The limits of detections were 55.7 pg/ml (0.95 pM) for H5N1 HA, 99.6 pg/ml (1.69 pM) for H7N9 HA, and 54.0 pg/ml (0.72 pM) for H9N2 HA antigens in phosphate buffered saline, and the sensors showed good selectivity and reproducibility. Such paper-based sensors are economical, flexible, robust, and easy-to-manufacture, with the ability to detect several avian influenza viruses.

Challenges and perspectives of multi-virus biosensing techniques: A review

“…Similarly, the ePAD approach can be adopted to enhance the flexibility of voltammetric biosensors for multiplexed virus detection [ 88 ]. A screen-printed carbon electrode on paper substrate immunosensors has recently proven its stability in detecting multiple avian influenza viruses [ 89 ]. The biosensor measured the peak current decrease using DPV, achieving 55.7 pg mL −1 , 99.6 pg mL −1 , and 54.0 pg mL −1 LOD for avian influenza virus (AIV) H5N1, H7N9 and H9N2 hemagglutinin antigens with 20–30 min antibody-antigen binding time and only 10 of samples.…”

“… Peak current decrease under DPV measurement. 2 h AIV H5N1, H7N9, H9N2 antigens PBS AIV H5N1: 55.7 pg mL −1 (0.95 pM) AIV H7N9: 99.6 pg mL −1 (1.69 pM) AIV H9N2: 54.0 pg mL −1 (0.72 pM) 20–30 min No [ 89 ] Antibody Single working electrode. Peak current decrease under DPV measurement.…”

Decadal Journey of CNT-Based Analytical Biosensing Platforms in the Detection of Human Viruses

“…Wang et al also reported that the aptamer could provide an LOD in the range of 10 fM, whereas the antibody showed an LOD of 1 pM in detecting the influenza A virus. Using flexible and stable SP-CNT-polydimethylsiloxane electrodes, a paper-based immunosensor for the label-free hemagglutinin antigen (HA) detection of several AIVs (H5N1, H7N9, and H9N2) was demonstrated by Lee et al [ 43 ], whereby immune responses were measured via DPV. The LOD values for the different viruses were 55.7 pg/mL for H5N1 HA, 99.6 pg/mL for H7N9 HA, and 54.0 pg/mL for H9N2 HA.…”

Development of disposable electrode for the detection of mosquito‐borne viruses

“…The electrochemical-based biosensor has recently been advanced, providing a high signal-to-noise in analyte detection. This is accomplished by the developing various nanomaterials as the signal transducer [25][26][27][28] with its signal amplification strategy. [29,30] However, the complex composition of the electrode, starting from the signal transducer, advanced nanomaterial, and bioconjugation, mostly fails to be adapted for POC diagnostic tools for on-site biosensor.…”