This study aimed to determine the prevalence of extended-spectrum beta-lactamases (ESBLs) in Enterobacteriaceae and to characterize the genetic composition of ESBL determinants among Enterobacteriaceae isolates from healthy people in Guangzhou, China. A total of 200 rectal swab samples were collected from healthy asymptomatic individuals and tested for ESBL production using ChromID ESBL agar. Phenotypic ESBL producers were screened for blaCTX-M, blaTEM, and blaSHV genes using PCR and DNA sequencing. The prevalence of ESBL-producing Enterobacteriaceae among rectal swab samples was 69.5%. All ESBL-producing isolates harbored blaCTX-M genes (n=138) except for one isolate that harbored blaSHV-2a. Eleven CTX-M ESBL genes were detected. The most predominant CTX-M-type genes were blaCTX-M-14 (n=82), followed by blaCTX-M-55 (n=19), blaCTX-M-65 (n=10), and blaCTX-M-27 (n=9). Isolates carrying blaCTX-M-38,-3,-15,-14b,-98,-121 and -123 were also identified. Molecular homology analysis of the selected isolates was performed by phylogenetic grouping and multilocus sequence typing and indicated that the predominant clone belonged to A-CC10. This study showed a high rate of CTX-M-type ESBL genes among Enterobacteriaceae isolates from healthy individuals in China.