A novel direct readout colorimetric optical glucose sensor strip was constructed based on a three-layer film, including a green-emitted CdTe/CdS quantum dots (QDs) layer as a stable color background, a redfluorescent platinum-porphyrin oxygen-sensing layer and a glucose oxidase layer. The sensor achieved high resolution (up to 0.2mmolL−1) glucose determination with a detection range from0 to 3.0mmolL−1. A “glucose ruler” which acts as a glucose standard colorimetric cardwas obtained. Glucose concentration could easily be directly readout using the “glucose ruler”, which made the glucose determination rapid, convenient and easy. The effects of pH, salinity and temperature were systematically investigated. The prepared sensor was finally applied for glucose sample analysis, compared with the “glucose ruler”, accurate results could be directly readout.