DNA biosensor based on polyaniline (PANI)-poly vinyl sulphonate (PVS) has been fabricated using electrochemical entrapment technique for detection of organophosphorus pesticides (chlorpyrifos and malathion). These double stranded calf thymus DNA (dsCT-DNA) entrapped PANI-PVS/indium-tin-oxide (ITO) bioelectrodes have been characterized using square wave voltammetry (SWV), Fourier transform infra-red spectroscopy (FT-IR), scanning electron microscopy (SEM) and electrochemical impedance techniques, respectively. These dsCT-DNA entrapped PANI-PVS/ITO bioelectrodes have been found to have response time of 30 s, stability of about 6 months and detection limit for chlorpyrifos and malathion as 0.5 ppb and 0.01 ppm, respectively. (c) 2008 Elsevier Ltd. All rights reserved.