A disposable glucose test strip with an integrated sensing/diffusion-limiting layer was developed. A formulation containing filler, glucose oxidase and electronic mediator was screen-printed over two carbon electrodes to form an integrated sensing/diffusion-limiting layer. On rehydration, the integrated layer does not break up, but swells to form a gelled and three-dimensional meshy reaction zone on the surface of the underlying conductive elements in which reactants and mediator move freely, but interfering species such as red blood cells containing oxygenated hemoglobin are excluded. On the same time, the integrated layer maintains a rate of permeation of the analyte through it with a variation of less than 10% at temperatures ranging from 15 degrees C to 42 degrees C. This biosensor is substantially insensitive to interferents and essentially independent to relevant temperature, which provides a more reliable reading of actual blood glucose value in human whole blood. (C) 2009 Elsevier Ltd. All rights reserved.