In this study, cationic carbon dots (CDs) were prepared from p-phenylenediamine (pPDA) via a one-step hydrothermal method and used to trigger the aggregation and dispersion of gold nanoparticles (AuNPs) for the colorimetric detection of nucleic acids. Physicochemical characterization results revealed that the CDs are enriched with positively charged surface functional groups with an average size of ∼11 nm. The interaction between the CDs and AuNPs was confirmed via fluorescence and absorption studies. Absorption spectroscopic results revealed that the primary surface plasmon resonance (SPR) band of the AuNPs decreased upon introduction of CDs, and a new band emerged at ∼600 nm, indicating the aggregated assembly of AuNPs. Upon the introduction of double-stranded deoxyribonucleic acid (DNA), the band corresponding to the aggregated AuNPs showed a continuous decrease, accompanied by a simultaneous increase in the primary SPR band, leading to a noticeable purple-to-red color transformation. Based on this phenomenon, a colorimetric assay for DNA was developed, which relies on the interaction between negatively charged DNA and cationic CDs, leaving the AuNPs dispersed. The assay exhibited a linear response within a DNA concentration range of 0.7–14 nM with a detection limit of 1.70 nM. Selectivity results showed that colorimetric assays are specific for both DNA and single-stranded DNA (ssDNA). Smartphone-assisted detection was developed by monitoring the colorimetric response of a AuNPs/CDs probe. As a proof-of-concept experiment, the AuNPs/CDs probe was used to visualize the loop-mediated isothermal amplification (LAMP) of Escherichia coli (E. coli), a robust indicator of sewage contamination in water.