The invention discloses a two-step biological leaching method for full-value recycling of valuable metals in a waste circuit board. The two-step biological leaching method comprises the following steps that S1, pretreatment is conducted, specifically, the waste circuit board is smashed and then sorted, and thus metal particles are obtained; S2, leaching of the non-ferrous metals is conducted, specifically, an ore-leaching microorganism is cultured in a culture medium containing Fe2+, so that Fe2+ is converted into Fe3+, then Fe3+ reacts with the metal particles in the step S1, and thus non-ferrous metal leachate and precious metal residues are obtained; S3, leaching of precious metals is conducted, specifically, the precious metal residues in the step S2 react with a precious metal leaching agent solution formed by the mode that lime water absorbs HCN, and thus precious metal leachate is obtained; and S4, the non-ferrous metal leachate in the step S2 and the precious metal leachate inthe step S3 are extracted and recycled correspondingly. By adopting a two-step leaching process, full-value extraction of the non-ferrous metals in the waste circuit board can be achieved, and an environment-friendly technological scheme is provided for recycling of the metals in the waste circuit board.