Rare sugars are defined as monosaccharides and their derivatives that rarely exist in nature, according to the International Society of Rare Sugars. d-Allose (3-epi d-glucose) is one of the rare sugars, for which various physiological activities have recently been found, with increasing attention to its applications to bio-industry. Until now, however, there is no convenient method of measuring these sugars in a specific manner. For detecting d-allose, three consecutive enzyme reactions were devised by fabricating of a reaction batch chamber packed with l-rhamnose isomerase (LRI), d-tagatose 3-epimerase (DTE) and a screen-printed electrode, on which d-fructose dehydrogenase (DFDH) was immobilized. To obtain a substantial sensing system, extensive experimental parameters were optimized. These included the concentration of photo-crosslinkable poly (vinyl alcohol) bearing stilbazolium groups (PVA-SbQ), reaction ratios, and temperature of the batch chamber. By adopting the three consecutive enzyme reactions, an undesirable reverse reaction was minimized. As a result, the developed sensor system exhibited a good linear response on d-allose in the range from 0.1 to 50mM (r 2 =0.998). The system has an apparent advantage over the previous chromatography approach in terms of simplicity and inexpensiveness.