<p>Antibiotic resistance genes (ARGs) are emerging pollutants that pose a serious threat to public health. Existing evidence indicates that cave environments are also reservoirs of ARGs, but relevant research has not received widespread attention. It is generally believed that even in karst cave environments with little human disturbance, there are diverse ARGs, whose distribution are jointly driven by specific environmental factors and microbial communities. However, this hypothesis has not been confirmed yet. Accordingly, the Shuanghe Cave in Guizhou Province, China was selected to analyze the physical and chemical parameters of the surface-layer mixtures inside the cave. Moreover, ARGs and microbial diversity, potential host bacteria of ARGs, as well as the influence of environmental factors on them were assayed and analyzed using metagenomic technology. Results indicated that 21 ARG types and 728 ARG subtypes were annotated according to the CARD database, with multidrug and <i>macB</i> being the predominant ones. High-risk mobile <i>optrA</i>, <i>tetA</i> and <i>mupA</i> genes were clearly identified. Correlation analysis between ARGs and microorganisms suggested that Proteobacteria and Actinobacteria might serve as key potential hosts and disseminators for multiple ARGs. Redundancy analysis (RDA) and Pearson correlation analysis further showed that SO₄²⁻ and NO₃⁻ were the key environmental factors influencing the composition and distribution of ARGs and microorganisms in the cave. This study confirmed the ecological risks of karst caves as ARGs reservoirs and also provided important insights into the distribution and transmission mechanisms of ARGs in caves.</p>