Enzymatic amplification detection of peanut allergen Ara h1 using a stem-loop DNA biosensor modified with a chitosan-mutiwalled carbon nanotube nanocomposite and spongy gold film.

In this paper, a highly sensitive biosensor was constructed for peanut allergen Ara h1 detection. The biosensor was constructed by coating a glassy carbon electrode with a chitosan-mutiwalled carbon nanotube nanocomposite and then adding a spongy gold film via electro-deposition to increase the effective area. The probe switched from an "on" to an "off" state in the presence of target DNA, which detached biotin from the electrode surface. This also detached streptavidin-horseradish peroxidase (HRP-SA), which was bound to the electrode via specific interaction with biotin. The HRP-SA catalyzed chemical oxidation of hydroquinone by H2O2 to form benzoquinone, and when it was detached, electrochemical reduction of the signal of benzoquinone could be used to monitor DNA hybridization via chronoamperometry. Under optimum conditions, a wide dynamic detection range (3.91 × 10(-17)-1.25 × 10(-15) mol L(-1)) and a low detection limit (1.3 × 10(-17) mol L(-1)) were achieved for the complementary sequence. Furthermore, the DNA biosensor exhibited an excellent ability to discriminate between a complementary target and a one-base mismatch or non-complementary sequence. The sensor was successfully applied to Ara h1 analysis in peanuts.