Background: In recent years, identifying players with injury risk through physical fitness assessment has become a hot topic in sports science research. Although practitioners have conducted many studies on the relationship between physical fitness and the likelihood of injury, the relationship between the two remains indeterminate. Consequently, this study utilized machine learning to preliminary investigate the relationship between individual physical fitness tests and injury risk, aiming to identify whether patterns of physical fitness change have an impact on injury risk. Methods: This study conducted a retrospective analysis by extracting the records of 17 young female basketball players from the sport-specific physical fitness monitoring and injury registration database in Fujian Province. Sports-specific physical fitness tests included physical performance, physiological, biochemical, and subjective perceived responses. The data for each player was standardized individually using Z-scores. Synthetic minority over-sampling techniques and edited nearest neighbor algorithms were used to sample the training set to address the negative impact of class imbalance on model performance. Feature extraction was performed on the dataset using linear discriminant analysis, and the prediction model was constructed using the cost-sensitive neural network. Results: The 10 replicate 5-fold stratified cross-validation showed that the lower limb non-contact injury prediction model based on the cost-sensitive neural network had achieved good discrimination and calibration (average Precision: 0.6360; average Recall: 0.8700; average F2-Score: 0.7980; average AUC: 0.8590; average Brier-score: 0.1020), which could be well applied in training practice. According to the attribution analysis, agility and speed were important physical attributes that affect youth female basketball players' non-contact lower limb injury risk. Specifically, there was enhance in the performance of the 1-min double under, accompanied by an increase in urinary ketone and urinary blood levels following the agility test. The 3/4 basketball court sprint performance improved, while urinary protein and RPE levels decreased after the speed test. Conclusion: The sport-specific physical fitness change pattern can impact the lower limb non-contact injury risk of young female basketball players in Fujian Province, specifically in terms of agility and speed. These findings will provide valuable insights for planning athletes' physical training programs, managing fatigue, and preventing injuries.