The Duocaima carbonate-hosted Pb-Zn deposit is a newly found large deposit in the southern area of Qinghai Province. In this paper, the characteristics, genesis, significance to Pb-Zn mineralization of the widely developed breccias, and the ore-forming process have been carefully studied based on geological documentation of drilling holes, microscopic observations of petrography and microstructure and some stable isotope measurements. Based on the compositions of the clast and matrix, the breccias can be classified into three types: limestone clasts cemented by marl; limestone clasts with fine-grained calcareous materials; and limestone clasts cemented by hydrothermal calcite. The mineralization in the first type of breccia is weak, whereas it is strong in the latter two types of breccias. According to the locations of occurrence and structural characteristics of the breccias along with the relationship between the breccias and mineralization, part of the limestone clasts that are cemented by marl and outcrop in the contact zone between the Wudaoliang Formation (Nw) and the underlying Jiushidaoban Formation (Pj) are attributed to synsedimentary fault-genetic breccia, whereas the last of the limestone clasts that are cemented by marl and developed in the Jiushidaoban Formation (Pj) are attributed to the breccia generated by karst cave collapse; the limestone clasts with fine-grained calcareous materials and the limestone clasts cemented by hydrothermal calcite are attributed to breccia formed by hydrothermal dissolution. The breccia formed by karst collapse had consistently evolved for a long period of time, while the breccias with other origins were formed around the period of mineralization (i.e., about or slightly later than 20–16 Ma). The breccia generated by karst cave collapse and hydrothermal dissolution are somewhat related; the formation of the breccia from karst cave collapse provided open space for the later mineralization and reaction between hydrothermal fluids and host rocks, and the subsequent strong dissolution by hydrothermal fluids transformed some of the breccia formed earlier by karst cave collapse. Meanwhile, carbonate host rocks with breccias and brecciaed mineralization can be a potential sign of Mississippi Valley Type (MVT) deposits and important indicators for regional mineral exploration. The δ13CV-PDB, δ18OV-SMOW, and 87Sr/86Sr values of hydrothermal calcite in the Duocaima deposit range from 4.3‰ to 7.1‰, 14.9‰ to 20.1‰, and 0.707494 to 0.708185, respectively; the δ13CV-PDB, δ18OV-SMOW, and 87Sr/86Sr values of the host limestones of the Jiushidaoban Formation range from 3.6‰ to 5.3‰, 18.0‰ to 20.5‰, and 0.707372 to 0.707945, respectively. The δ13CV-PDB and 87Sr/86Sr values of hydrothermal calcite and limestone are similar, indicating single sources of C and Sr in this deposit, with the likely source being the limestone of the Jiushidaoban Formation. The minor scattering of the δ18OV-SMOW values suggests that different O isotope fluids underwent the isotope exchange reaction. The C-O-Sr isotope characteristics indicate that the host limestones experienced a dissolution and precipitation process during mineralization, which is beneficial to improving the porosity of host rocks and promoting the precipitation of metal sulfides. The δ34SV-CDT value of the breccia-type mineralization sulfides ranges from ?30.4‰ to ?0.3‰; that is, the δ34SV-CDT value is negative with considerable variation, illustrating that during the breccia-type mineralization process, the bacteriogenic reduction of sulfates provided the vast majority of sulfur, whereas the thermochemical reduction of sulfates was relatively unimportant. The brecciation that occurred as a result of karst cave collapse was mainly generated by the dissolution of groundwater; however, the brecciation related to hydrothermal dissolution and mineralization processes were caused by mixing of different fluids.