BACKGROUND: Predicting associations between microbes and diseases is crucial for clinical diagnosis and therapy. However, biological experiments are time-intensive, necessitating efficient computational models. Traditional models rely on existing microbe-disease associations, but limited data often restricts their effectiveness. This scarcity of information hinders the construction of a comprehensive association network, prompting the need for innovative solutions. METHODS: We propose RKGATMDA, a deep learning framework for microbe-disease association prediction. Utilizing a graph attention network, RKGATMDA learns representations from the microbe-disease association network. To address the limitation of insufficient association information, we introduce Random K-Nearest Neighbors to uncover latent relationships, enhancing representation learning. During each training iteration, associations are expanded based on attention scores, and a multi-head attention mechanism integrates diverse features, enabling RKGATMDA to capture comprehensive interactions between microbes and diseases. RESULTS: Results Experimental results show that RKGATMDA achieves AUC values of 0.8906 in 5-fold cross-validation, 0.8999 in global leave-one-out cross-validation, and 0.7246 in local leave-one-out cross-validation, outperforming previous methods such as ABHMDA, KATZHMDA, LRLSHMDA, BiRWHMDA, and NTSHMDA. Case studies on asthma, colon cancer, and colorectal carcinoma further validate its predictive power. CONCLUSION: Our findings demonstrate that RKGATMDA effectively predicts microbe-disease associations, with at least 9 out of the top 10 prediction pairs validated by biological evidence. This highlights the potential of RKGATMDA as a valuable tool in microbial-disease research, offering a promising approach for identifying novel associations and advancing our understanding of microbial pathogenesis.