Abstract: Abstract: Blueberry is one of the most economically important nutritional fruits, particularly in popularity with the remarkable blue color and high health benefits of antioxidant potential. The purpose of this study was to establish a comprehensive quality evaluation of highbush blueberry using principal component analysis. A total of 11 quality indicators were selected (including single fruit weight, diameter of fruit, longitudinal diameter of fruit, fruit shape index, fruit hardness, soluble solids mass fraction, soluble sugar mass fraction, titration acidity mass fraction, sugar acid ratio, vitamin C mass fraction, and anthocyanin mass fraction) for 26 highbush blueberry cultivars. There was a significant difference in the variation coefficient of the 11 fruit quality index of blueberry cultivars. In terms of fruit quality indexes, the coefficients of variation were 33.67%, 32.25%, 30.84%, and 28.48% for the sugar-acid ratio, titratable acidity, single fruit weight, and anthocyanin mass fraction, respectively. There was only 10.00% or less in the coefficient of variation of the fruit shape index. The medium variation coefficient (10.00% to 20.00%) was recorded in the six remaining fruit quality indices. Besides, principal component analysis was used to extract four principal components with eigenvalues greater than 1, where the cumulative variance contribution rate was 78.19%. Specifically, the contribution rate of the first principal component (28.05%) was determined as the reflected fruit size, i.e., fruit diameter, single fruit weight, and fruit longitudinal diameter. The contribution rate of the second principal component (25.89%) was determined as the parameters related to the sweet taste and nutritional quality of fruit, such as the soluble solid mass fraction, soluble sugar mass fraction, sugar-acid ratio, anthocyanin mass fraction, and vitamin C mass fraction. The third principal component accounted for 14.73% of the fruit acidity and fruit firmness, in terms of the titratable acidity and fruit firmness. The contribution rate of the fourth principal component was 9.52%, which was determined by the fruit shape index. Furthermore, six of 11 evaluated indexes (single fruit weight, fruit shape index, soluble sugar mass fraction, titratable acidity mass fraction, anthocyanin mass fraction, and fruit firmness) were screened as the evaluation indexes of blueberry quality, which were characterized by the fruit size, fruit shape, sweet, sour, nutritional, and texture index. An excellent level order of 26 blueberry cultivars was achieved in the quality evaluation model: Big bluegold, Legacy, Duke, Sunrise, Weymouth, Bluechip, Earliblue, Bluecrop, Patriot, and Bonus. The findings can provide a theoretical reference to evaluate the blueberry quality, and then screen the blueberry varieties with excellent quality, particularly for the rational development and application of blueberry cultivars.