This study focuses on the fabrication of a novel, flexible and disposable textile based biosensing platform by the use of an absorbent microfibrous nonwoven substrate as the base material. This platform was fabricated via photolithography technique. Physical barriers were designed using a hydrophobic photo-resist polymer which defined the liquid penetration pathways on the fabric surface. A good hydrophilic/hydrophobic contrast of the fabricated patterns on the fabric and a well-controlled liquid capillary penetration was achieved in the patterns. The potential of the system was tested by constructing anenzyme biosensor based on colorimetric detection of hydrogen peroxide. To obtain a more enhanced and reproducible signal, the reservoirs were modified with gelatin and a linear working range of 0.1-0.6 mu M H2O2 was obtained. The system could work on temperatures as high as 50 degrees C without any loss in the signal and in a pH range of 3.0-7.0. This bio-sensing platform may later be combined by H2O2 producing oxidases such as glucose oxidase, lactate oxidase, etc. and used for the rapid detection of various kinds of important analytes. (C) 2014 Elsevier B.V. All rights reserved.