Abstract
An improved ELISA for the determination of tobacco mosaic virus (TMV) with linear sweep voltammetry based on a new system of p-aminophenol (PAP)- H 2 O 2 -horseradish peroxidase (HRP) has been developed. The enzymatic product 3-[(4-hydroxyphenyl)amino]-4-(2-amino-5-hydroxyphenyl)-6-[(4-hydroxyphenyl)imino]-2,4-cyclo-hexadiene-1-one, produced from HRP catalyzing the oxidation of PAP with H 2 O 2 , yields a sensitive linear sweep voltammetric response at a potential of -0.45 V (vs. SCE) in Britton-Robinson (BR) buffer solution. By using this voltammetric peak, HRP can be measured with a detection limit of 0.4 mU/L and a linear range of 1.0 ∼ 1.0 × 10 2 mU/ L. The detection limit for the clarified TMV is 4.0 ng/mL and the highest dilution ratio detected for the infected leaf sap is 1:3.9 × 10 6 . The processes of the enzyme-catalyzed reaction and the electro-reduction of the product of the enzyme-catalyzed reaction have been investigated.
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