Nanopore-Based\nBiomimetic ssDNA Reporters for Multiplexed\nDetection of Meat Adulteration

Meat adulteration detection is crucial for ensuring food\nsafety,\nprotecting consumer rights, and maintaining market integrity. However,\nthe current methods face challenges in achieving multiplexed detection\nthrough efficient signal conversion and output. This study introduces\na nanopore-based approach for the simultaneous detection of multiple\nmeat adulteration. Leveraging the interaction between <sup>D</sup>A<sup>D</sup>A dipeptide and vancomycin, we designed a series of\nbiomimetic ssDNA reporters with <sup>D</sup>A<sup>D</sup>A-Van tags\nfor multiplexed signal output. These ssDNA reporters can generate\nhighly distinctive current blockage signals, which can be discriminated\nsimultaneously by the current waveform. Combined with PCR-based strand\ndisplacement, ssDNA reporters can be released from magnetic beads\nin the presence of the target gene and are analyzed using α-hemolysin\n(α-HL) nanopores for multiplexed signal output. The signal frequency\nfor each target gene has a linear relationship with the specific concentration\nrange (for duck: 1 × 10<sup>–3</sup> ∼ 10 nmol/L,\nfor pork: 1 × 10<sup>–2</sup> ∼ 10 nmol/L, and\nfor chicken: 1 × 10<sup>–3</sup> ∼ 10 nmol/L).\nThe limits of detection are as follows: 0.418 pmol/L for duck, 4.473\npmol/L for pork, and 0.531 pmol/L for chicken. This method effectively\nenables the simultaneous detection of adulterated chicken, pork, and\nduck meat in lamb samples and holds potential for broad applications\nin ensuring the authenticity and safety of meat products.