We herein developed a novel colorimetric polydiacetylene (PDA) sensor for very convenient detection of clinical DNA samples based on the interaction between an intercalator and dsDNA. We modified the terminal carboxyl group of a diacetylene monomer (10,12-pentacosadiynoic acid; PCDA) with the intercalator 9-aminoacridine (9AA) and prepared 9AA-modified PDA liposomes containing PCDA-9AA/PCDA/phospholipid (1,2-dimyristoyl-rac-glycero-3-phosphocholine) at a molar ratio of 1.5:6.5:2.0. The PDA sensor underwent an obvious color transition from blue to red in the presence of dsDNA molecules that were PCR-amplified from genomic DNA due to the insertion of the 9AA head group of PDA into the dsDNA. DNA concentrations as low as 20 nM and relatively small molecules (around 100 base pairs) could be detected by the sensor within 1 h without DNA electrophoresis. This novel colorimetric method is simple, does not require any instrument, and is therefore appropriate for POCT or portable molecular diagnostic kit. (C) 2015 Elsevier B.V. All rights reserved.