Research Article Nanobioprobe for the Determination of Pork Adulteration in

which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. We report the development of a swine-specific hybrid nanobioprobe through a covalent integration of a fluorophore-labeled 27-nucleotide AluI-fragment of swine cytochrome b gene to a 3 nm gold nanoparticle for the determination of pork adulteration in processed meat products. We tested the probe to estimate adulterated pork in ready-to-eat pork-spiked beef burgers. The probe quantitatively detected 1–100 % spiked pork in burger formulations with ≥90 % accuracy. A plot of observed fluorescence against the known concentration of AluI-digested pork DNA targets generated a concave curve, demonstrating a power relationship (y = 2.956x0.509) with a regression coefficient (R2) of 0.986. No cross-species detection was found in a standard set of pork, beef, chicken, mutton, and chevon burgers. The method is suitable for the determination of very short-length nucleic acid targets which cannot be estimated by conventional and real-time PCR but are essential for the determination of microRNA in biodiagnostics and degraded DNA in forensic testing and food analysis. 1