In dynamic scenarios, the status of a Radio Frequency Identification (RFID) system fluctuates with environmental changes. The key to improving system efficiency lies in the real-time monitoring and evaluation of the system status, along with adaptive adjustments to the system parameters and read algorithms. This paper focuses on the status changes in RFID systems in dynamic scenarios, aiming to enhance system robustness and reading performance, ensuring high link quality, reasonable resource scheduling, and real-time status evaluation under varying conditions. This paper comprehensively considers the system parameter settings in dynamic scenarios, integrating the interaction model between readers and tags. The system’s real-time status is evaluated from both the physical layer and the Medium Access Control (MAC) layer perspectives. For the physical layer, a link quality evaluation model based on Uniform Manifold Approximation and Projection (UMAP) and K-Means clustering is proposed from the link quality. For the MAC layer, a multi-criteria decision-making evaluation model based on combined weighting and the Technique for Order Preference by Similarity to Ideal Solution (TOPSIS) is proposed, which comprehensively considers both subjective and objective factors, utilizing the TOPSIS algorithm for an accurate evaluation of the MAC layer system status. For the RFID system, this paper proposes a real-time status evaluation model based on the Classification and Regression Tree (CART), which synthesizes the evaluation results of the physical layer and MAC layer. Finally, engineering tests and verification were conducted on the RFID robot system in mobile scenarios. The results showed that the clustering average silhouette coefficient of the physical layer link quality evaluation model based on K-Means was 0.70184, indicating a relatively good clustering effect. The system status evaluation model of the MAC layer, based on the combined weighting-TOPSIS method, demonstrated good flexibility and generalization. The real-time status evaluation model of the RFID system, based on CART, achieved a classification accuracy of 98.3%, with an algorithm runtime of 0.003 s. Compared with other algorithms, it had a higher classification accuracy and shorter runtime, making it well suited for the real-time evaluation of the RFID robot system’s status in dynamic scenarios.