Abstract: Objective To investigate the distribution of enterovirus (EV) serotypes of 759 cases in Guangzhou during 2014-2015. Methods Rectal swabs were collected from children with suspected infection of enterovirus and detected for pan-EV-, EV71- and Coxsackie virus A group 16(CA16)-specific nucleic acid using real-time reverse transcription-polymerase chain reaction (RT-PCR). The serotype of EV-positive samples was identified using semi-nested RT-PCR and sequencing for VP1 gene product. Results A total of 759 (71.6 %) were detected EV positive among the 1 060 specimens during 2014-2015. Eleven serotypes were identified circulating in both of years. The top five predominant serotypes were CA16, EV71, CA4, CA10, CB5 in 2014 and CA6, EV71, CA10, CA16 and CA2 in 2015 respectively. CA16 and EV71 accounted for 77.1% in 2014, contrasting that CA6 accounted for 51.7% in 2015. The sex ratio ranged from 1.8 to 5.3. Children under 3 years old were more likely suffering from infection (60.1%, 456/759). EV could be detected throughout the year with CA16, EV71, CA10 and CA2 peaked in May while CA6 in July. Clinical manifestation was mild in most infection cases. However, severe cases were observed in 12.7% of EV71 infection and occasionally in CA16, CA10, CA4 and CA6 infections. Conclusion Eleven serotypes of EV, with different spectrum, circulated in Guangzhou during 2014-2015. Clinical manifestation in most EV infection were mild.