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In the present study, novel molecularly imprinted polymer (MIP) nanocomposite films were prepared for the detection of porcine double-stranded deoxyribonucleic acid (dsDNA) in food samples. A fragment of dsDNA from the pork mitochondrial genome was chosen as a template molecule for porcine identification. The molecularly imprinted polymer (MIP) nanocomposite films of polypyrrole-TiO2(PPy/TiO2) were synthesized electrochemically on the surface of indium-tin-oxide (ITO) glass electrode. A strong voltammetric (DPV) response was observed for capturing dsDNA from the samples containing porcine. The batch rebinding experiments revealed that PPy/TiO2MIP possessed excellent binding capacity (Q= 87.44%) and a high imprint factor (2.30) than non-imprinted polymers (NIPs). The selectivity experiments indicated that MIP had excellent recognition selectivity for porcine dsDNA. MIP exhibited higher relative selectivity coefficient (K′ = 4.26 µM/µL) toward porcine dsDNA as compared to structural analogue beef dsDNA (K′ = 0.23 µM/µL) and chicken dsDNA (K′ = 0.21 µM/µL). This could be attributed to the introduction of the imprinted cavities exactly matching the porcine dsDNA on the surface of MIP nanocomposite film. Furthermore, PPy/TiO2MIP showed a high reusability and low LOD (2.06 μM) and LOQ (6.89 μM) values.