Preparation of a Biosensor for DNA Damage Detection and its Application in Rapid Detection of Aflatoxin B1 in Feed
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Abstract:
In this study, a single-stranded DNA/Au (ssDNA/Au) electrode was prepared by aligning a self-assembled monolayer of thiolated ssDNA onto an Au electrode surface. Under optimal conditions, ssDNA/Au hybridized with the complementary DNA (cDNA) fragment to produce a double-stranded DNA/Au (dsDNA/Au) electrode, which was a useful and cost-effective electrochemical DNA biosensor for the rapid detection of aflatoxin B1 (AFB1). Electrochemical methods such as cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were applied to characterize the electrochemical behavior of ferricyanide and methylene blue (MB) (as electron mediators) on the surface of ssDNA/Au and dsDNA/Au electrodes. A certain concentration of AFB1 reacted with dsDNA/Au to induce DNA damage, thereby reducing the electrochemical signals from MB and allowing for rapid detection of AFB1. Under optimized conditions (ssDNA was self-assembled on gold electrodes for 14 h at 4 ℃ and hybridized with cDNA at 37 ℃ for 2 h to produce the dsDNA/Au electrode; AFB1 solution induced DNA damage for 22 min at 37 ℃), the linear detection range for AFB1 was 10~500 ng/mL, with a satisfactory recovery ranging from 95.99% to 104.57%. This method of AFB1-induced DNA damage detection could be explored as a useful, quantitative method for accurate and rapid detection of AFB1.
