Multilayer graphene–gold nanocomposite modified stem-loop DNA biosensor for peanut allergen-Ara h1 detection

In this study, we developed an electrochemically-amplified, stem-loop DNA biosensor to detect the peanut allergen Ara h1. Specifically, we electrodeposited a multilayer graphene–gold nanocomposite onto a glassy carbon electrode and then immobilised a thiolated hairpin DNA–biotin probe onto the modified electrode surface. The multilayer graphene–gold composite has good dispersion ability, and can amplify the electrochemical signal due to its high electron-transfer efficiency. The probe was switched to an “off” state in the presence of target DNA. The prepared biosensor demonstrated a linear response ranging from 10−16 to 10−13 M, with an ultrasensitive detection limit of 0.041 fM. Moreover, the biosensor showed excellent selectivity, as well as the ability to discriminate between a complementary target and a one-base mismatch or non-complementary sequence. Results show that this prepared DNA biosensor can be successfully used to detect the peanut allergen Ara h1 in a peanut milk beverage. Findings can be applied to the prevention of allergic reactions, thus improving human health and safety.