Healthcare-associated infection (HAI) and antimicrobial resistance (AMR) are major public health threats. The aims of this research were to estimate HAI prevalence, AMR incidence, as well as to assess adoption and implementation of infection prevention and control (IPC) in Mainland China. In 2015, the pooled HAIs prevalence of all hospitals in Dongguan was 2.9%. Between 2006 and 2016, the weighted HAI prevalence in Mainland China, as identified in the systematic review, was 3.1%. Incidence proportion of AMR in Dongguan was lower than average incidence proportion in mainland China. Most of the ECDC key and WHO core components were reported to be implemented by acute care hospitals in China, but significant gaps in effective IPC were identified. This is the first multilevel research on HAI and AMR in the context of implementing key elements for effective IPC. These outcomes serve as a reference for future IPC strategies in Mainland China.