The study aims were to present in vitro susceptibilities of clinical isolates from Gram-negative bacteria bloodstream infections (GNBSI) collected in China. GNBSI isolates were collected from 18 tertiary hospitals in 7 regions of China from 2018 to 2020. Minimum inhibitory concentrations were assessed using a Trek Diagnostic System. Susceptibility was determined using CLSI broth microdilution, and breakpoints were interpreted using CLSI M100 (2021). A total of 1,815 GNBSI strains were collected, with E. coli (42.4%) and Klebsiella pneumoniae (28.6%) being the most prevalent species, followed by P. aeruginosa (6.7%). Susceptibility analyses revealed low susceptibilities (<40%) of ESBL-producing E. coli and K. pneumonia to third-/fourth-generation cephalosporins, monobactamases, and fluoroquinolones. High susceptibilities to colistin (95.0%) and amikacin (81.3%) were found for K. pneumoniae, while Acinetobacter baumannii exhibited a high susceptibility (99.2%) to colistin but a low susceptibility to other antimicrobials (<27.5%). Isolates from ICUs displayed lower drug susceptibility rates of K. pneumoniae and A. baumannii than isolates from non-ICUs (all P < 0.05). Carbapenem-resistant and ESBL-producing K. pneumoniae detection was different across regions (both P < 0.05). E. coli and K. pneumoniae were major contributors to GNBSI, while A. baumannii exhibited severe drug resistance in isolates obtained from ICU departments.