In this Communication, we report on the first preparation of conjugation polymer poly(2,3-diaminonaphthalene) (PDAN) microspheres via chemical oxidation polymerization of 2,3-diaminonaphthalene (DAN) monomers by ammonium persulfate (APS) at room temperature. We further demonstrate the use of PDAN microspheres as a novel quencher for fluorescence-enhanced nucleic acid detection.