ABSTRACTCarbapenem-resistantKlebsiella pneumoniae(CRKP) has disseminated globally and is difficult to treat, causing increased morbidity and mortality rates in critically ill patients. We conducted a multicenter cross-sectional study of intensive care unit (ICU) inpatients in 78 hospitals to investigate the prevalence and molecular characteristics of CRKP in Henan Province, China, a hyperepidemic region. A total of 327 isolates were collected and downsampled to 189 for whole-genome sequencing. Molecular typing revealed that sequence type 11 (ST11) of clonal group 258 (CG258) was predominant (88.9%,n= 168), followed by ST2237 (5.8%,n= 11) and ST15 (2.6%,n= 5). We used core genome multilocus sequence typing (cgMLST) to further classified the population into 13 subtypes. Capsule polysaccharide (K-antigen) and lipopolysaccharide (LPS; O-antigen) typing revealed that K64 (48.1%,n= 91) and O2a (49.2%,n= 93) were the most common. We studied isolates collected from both the airway and the gut of the same patients and showed that intestinal carriage was associated with respiratory colonization (odds ratio = 10.80,P< 0.0001). Most isolates (95.2%,n= 180) showed multiple drug resistance (MDR), while 59.8% (n= 113) exhibited extensive drug resistance (XDR), and all isolates harbored eitherblaKPC-2(98.9%,n= 187) orblaCTX-MandblaSHVextended-spectrum beta-lactamases (ESBLs) (75.7%,n= 143). However, most were susceptible to ceftazidime-avibactam (CZA) (94.7%,n= 179) and colistin (97.9%,n= 185). We foundmgrBtruncations in isolates conferring resistance to colistin and mutations inblaSHVand OmpK35 and OmpK36 osmoporins in CZA-resistant isolates. Using a regularized regression model, we found that the aerobactin sequence type and the salmochelin sequence type, among others, were predictors of the hypermucoviscosity phenotype.IMPORTANCEIn this study, we address the ongoing epidemic of carbapenem-resistantKlebsiella pneumoniae, a critical threat to public health. The alarming genotypic and phenotypic convergence of multidrug resistance and virulence highlights the increasingly aggravated threat posed byK. pneumoniae. This calls for a combined effort of physicians and scientists to study the potential mechanisms and establish guidelines for antimicrobial therapies and interventions. To this end, we have conducted a genomic epidemiology and characterization study using isolates collected in a coordinated effort of multiple hospitals. Innovative biological discoveries of clinical importance are made and brought to the attention of clinical researchers and practitioners. This study presents an important advance in the application of genomics and statistics to recognize, understand, and control an infectious disease of concern.