To explore the effects of slope position and soil horizon on soil microbial biomass and abundance, chloroform fumigation extraction methods and real-time fluorescence-based quantitative PCR (Real-time PCR) were adopted to quantify the changes of soil microbial biomass C, N and abundance of bacteria and fungi, respectively. Soil samples were harvested from three horizons along profile, i. e., leaching horizon (A, 0-10 cm), transitional horizon (AB, 30-50 cm) and alluvial horizon (B, 70-100 cm), which were collected from the upper, middle and lower slope positions of a karst primary forest ecosystem. The results showed that slope position, soil horizon and their interaction significantly influenced the soil microbial biomass and abundance (P < 0.05). Different from A horizon, where SMBC was greater in lower than in upper slope position (P < 0.05), SMBC in AB and B horizons were highest in middle slope position. Similarly, SMBN was greater in lower than in upper slope position for A, AB and B horizons. Besides soil bacterial abundance in B horizon and fungal abundance in AB layer, the middle slope position had the highest value for all the three soil horizons (P < 0.05). Stepwise regression analysis showed that soil organic carbon, available nitrogen and pH were the key factors responsible for SMBC and SMBN variation, respectively, while the important factors responsible for the variation of bacteria abundance were available nitrogen and available phosphorus, and that for fungi abundance variation were available potassium.