Detection of TMV with ODA-H(2)O(2)-HRP voltammetric enzyme-linked immunoassay system.
o-Dianisidine (ODA)-H(2)O(2)-horseradish peroxidase (HRP) voltammetric enzyme-linked immunoassay system has firstly been used for the detection of tobacco mosaic virus (TMV). HRP catalyzes strongly the oxidation reaction of ODA by H(2)O(2), the product of which produces a sensitive second order derivative linear sweep voltammetric peak at potential of -0.56 V (versus SCE) in Britton-Robinson (BR) buffer. HRP activity has been measured with this voltammetric peak and TMV detected through immunoreaction. The detection limit for HRP is 9.25x10(-7) mU l(-1) and the linear range is 2.5x10(-6)-5.0x10(-4) mU l(-1). The detection limit for the clarified TMV is 0.25 ng ml(-1) and the highest dilution ratio detected for the infected leaf sap is 1:8x10(5). The sensitivity for TMV detection with this method is higher than that with the enzyme-linked immunosorbent spectrophotometric assay (ELISA) using ODA-H(2)O(2)-HRP system. The processes of the enzyme-catalyzed reaction and the electro-reduction of the product of the enzyme-catalyzed reaction have been described.
Jiao
K
K
Department of Applied Chemistry, Qingdao Institute of Chemical Technology, 53 Zhengzhou Road, 266042 Qingdao, People's Republic of China.
Zhang
S
S
Wei
L
L
Liu
C
C
Zhang
C
C
Zhang
Z
Z
Liu
J
J
Wei
P
P