Nowadays, the presence of highly toxic and persistent residues of pesticides in water and food around the world is becoming a serious problem, and so their rapid and sensitive detection is critical to human health. In this work, a 3D composite nanoparticle of porous PDA (polydopamine) microspheres and Au NPs (PPDA@Au NPs) was proposed as a SERS substrate to detect pesticides. Porous PDA as a substrate was first synthesized with F127 (Pluronic F127), dopamine hydrochloride, and 1,3,5-TMB (1,3,5-trimethylbenzene) under weakly alkaline conditions by a one-step method. Then, HAuCl4 was in situ reduced in the pores of PPDA spheres and grew sequentially for effecting the reducibility of PPDA. As a result, uniform 3D PPDA@Au NPs with "hot spots" were successfully synthesized as SERS substrates, which could effectively avoid the agglomeration of gold nanoparticles to greatly improve the sensitivity and uniformity of the SERS platform. At the same time, methyl parathion, 4-chlorophenol, and 2,4-D as representatives of pesticides were detected with the proposed PPDA@Au NP-based SERS platform, with detection limits lower than 7.26, 7.52, and 11 ng mL-1, separately. The current work presents a simple preparation method to prepare sensitive and uniform SERS platform PPDA@Au NPs, which have potential for applications in actual pesticide and drug testing.