In this work, the glassy carbon electrode modified by alizarin (AR) hybridized with graphene (CRG) and chitosan (CS) was prepared. The AR/CRG-CS electrode exhibited a pair of well-defined reversible redox peaks in 0.1 M phosphate buffer solution (PBS, pH 7.4) with the oxidation peak at −0.573 V and the reduction peak at −0.652 V. It exhibited a two-proton and two-electron redox process and a fast heterogeneous electron transfer rate with the rate constant (K s) of 1.69 s−1 using the cyclic voltammetry. Based on the characteristics of alizarin with the decreased current and negative potential shift after binding with human telomere DNA, a sensitive, simple, reliable and stable electrochemical method for the detection of human telomere DNA using the AR/CRG-CS electrode was developed. The electrochemical response showed a linear relationship with DNA concentration in the range of 8.0 × 10−8 −1.4 × 10−5 M (R = 0.9995) with the linear regression equation as ΔI pc (μA) = 8.4860 + 0.5366C (μmol) and the detection limit (S/N) as 2.0 × 10−8 M.