A Gold Nanoparticle-DNA Bioconjugate-Based Electrochemical Biosensor for Detection of Sus scrofa mtDNA in Raw and Processed Meat

The presence of pork content in food products can be a serious issue over an adulteration of food, particularly for consumers who pay attention to halal certification or other religious beliefs concerning the consumption of pork. Due to this fact, finding a method for the detection of pork content in food products that is sensitive, fast, and accurate is of interest. In this study, an electrochemical DNA biosensor with gold nanoparticle-DNA probe bioconjugates was developed to detect Sus scrofa mitochondrial DNA (mtDNA) using a gold-modified screen-printed carbon electrode (SPCE-Gold). The bioconjugates were formed by attaching the DNA probe with gold nanoparticles (AuNPs). Detection was carried out based on the optimum conditions resulting from the Box-Behnken experimental design that were 40 mu L of 153 mu g/mL bioconjugates; bioconjugate immobilization time, 20 min; and DNA hybridization time, 60 min. The hybridization of target synthetic DNA to the DNA probe was characterized by voltammetry based on the methylene blue indicator signal at a potential of about - 0.35 V. The results showed that the detection limit was 0.58 mu g/mL and the recovery was 101.74%. mtDNA samples from various raw and processed meat were isolated and cut with restriction enzyme Sal1. Hybridization of mtDNA samples to the gold nanoparticle-DNA probe bioconjugates anchored to the SPCE-Gold surface was characterized based on the current response generated by methylene blue as an indicator. The result shows that there is an increasing trend in the response of raw and processed meat samples containing pork DNA. The development of an electrochemical DNA biosensor, based on gold nanoparticle-DNA probe bioconjugates, has not been reported previously. This biosensor is selective towards 10% of the pork DNA content in the mixture. Therefore, this biosensor method can be used as an alternative method to distinguish samples containing pork, according to the detection limit.