TY - JOUR
T1 - Screen printed carbon electrode based electrochemical immunosensor for the detection of dengue NS1 antigen
AU - Parkash, Om
AU - Yean, Chan Yean
AU - Shueb, Rafidah Hanim
N1 - Publisher Copyright:
© 2014 by the authors.
PY - 2014
Y1 - 2014
N2 - An electrochemical immunosensor modified with the streptavidin/biotin system on screen printed carbon electrodes (SPCEs) for the detection of the dengue NS1 antigen was developed in this study. Monoclonal anti-NS1 capture antibody was immobilized on streptavidin-modified SPCEs to increase the sensitivity of the assay. Subsequently, a direct sandwich enzyme linked immunosorbent assay (ELISA) format was developed and optimized. An anti-NS1 detection antibody conjugated with horseradish peroxidase enzyme (HRP) and 3,3,5,5'-tetramethybezidine dihydrochloride (TMB/H2O2) was used as an enzyme mediator. Electrochemical detection was conducted using the chronoamperometric technique, and electrochemical responses were generated at -200 mV reduction potential. The calibration curve of the immunosensor showed a linear response between 0.5 μg/mL and 2 μg/mL and a detection limit of 0.03 μg/mL. Incorporation of a streptavidin/biotin system resulted in a well-oriented antibody immobilization of the capture antibody and consequently enhanced the sensitivity of the assay. In conclusion, this immunosensor is a promising technology for the rapid and convenient detection of acute dengue infection in real serum samples.
AB - An electrochemical immunosensor modified with the streptavidin/biotin system on screen printed carbon electrodes (SPCEs) for the detection of the dengue NS1 antigen was developed in this study. Monoclonal anti-NS1 capture antibody was immobilized on streptavidin-modified SPCEs to increase the sensitivity of the assay. Subsequently, a direct sandwich enzyme linked immunosorbent assay (ELISA) format was developed and optimized. An anti-NS1 detection antibody conjugated with horseradish peroxidase enzyme (HRP) and 3,3,5,5'-tetramethybezidine dihydrochloride (TMB/H2O2) was used as an enzyme mediator. Electrochemical detection was conducted using the chronoamperometric technique, and electrochemical responses were generated at -200 mV reduction potential. The calibration curve of the immunosensor showed a linear response between 0.5 μg/mL and 2 μg/mL and a detection limit of 0.03 μg/mL. Incorporation of a streptavidin/biotin system resulted in a well-oriented antibody immobilization of the capture antibody and consequently enhanced the sensitivity of the assay. In conclusion, this immunosensor is a promising technology for the rapid and convenient detection of acute dengue infection in real serum samples.
KW - Dengue diagnosis
KW - Electrochemical immunosensor
KW - NS1
KW - Screen printed carbon electrode
KW - Streptavidin/biotin
UR - http://www.scopus.com/inward/record.url?scp=84943263808&partnerID=8YFLogxK
U2 - 10.3390/diagnostics4040165
DO - 10.3390/diagnostics4040165
M3 - Article
AN - SCOPUS:84943263808
SN - 2075-4418
VL - 4
SP - 165
EP - 180
JO - Diagnostics
JF - Diagnostics
IS - 4
ER -