A hybridization chain reaction coupled with gold nanoparticles for allergen gene detection in peanut, soybean and sesame DNAs

Abstract

Food allergy is an abnormal immune response of the immune system to some foods, which has caused great harm to people's health. Therefore, it is particularly important to detect allergens in food. In this article, a hybridization chain reaction (HCR) was coupled with gold nanoparticles (AuNPs) to detect the allergen genes of peanut, soybean and sesame DNAs. Two hairpin probes (H1 and H2) were designed for the allergen target genes of peanut, soybean and sesame DNAs. In the presence of target DNA, the hybridization chain reaction was triggered by it producing long double-stranded DNA (dsDNA) products. In the gold nanoparticle system, long dsDNA couldn't be adsorbed on the surface of AuNPs. When the concentration of salt ions in the solution increased, gold nanoparticles accumulated and led to a decrease of ultraviolet absorption. In the absence of target DNA, no hybridization chain reaction occurred. The hairpin probes could be adsorbed on the surface of AuNPs and no accumulation happened for gold nanoparticles even if the concentration of salt ions in the solution was increased. This method required no enzymes and had a strong specificity, so it was very easy to distinguish target DNA from non-target DNA. The detection limit of three allergens detected by this method was as low as 0.5 nM. The feasibility of this method for the detection of commercial commodities had been demonstrated by the successful detection of the DNAs extracted from commercial commodities, which were treated with extreme thermostable single-stranded binding protein (ET SSB).