Diabetes mellitus is a prevalent chronic disease globally and detecting and monitoring diabetes progression typically involves assessing biomarkers such as fasting or postprandial blood glucose levels, glycated albumin, glycated hemoglobin, and 1,5-anhydroglucitol. In this study, a lateral flow assay (LFA) was developed to concurrently detect glucose levels and glycation ratios of human serum albumin. Ki et al. reported the specific enzymatic reactions and immunoassays, spiked glucose solutions, total human serum albumin, and glycated albumin were simultaneously measured. The developed sensor's performance was tested using clinical serum samples obtained from healthy subjects and diabetic patients. Reasonable correlations were observed between the glucose level and glycation ratios of clinical samples, with R-squared values of 0.932 and 0.930, respectively. The detection recoveries of the sensor were 85.80% for glucose and 98.32%