Abstract： Objective:To analyze the changes in the composition of the pathogens for hand, foot, and mouth disease (HFMD) in Zibo city, Shandong province from 2014 to 2022, and aware the epidemic patterns of the main pathogens of HFMD in this city, so as to provide suggestions and reference basis for the prevention and control of HFMD.Methods:Real time fluorescent reverse transcription PCR (RT-PCR) was used to detect enterovirus nucleic acid in 4 053 pharyngeal swab samples from Zibo city from 2014 to 2022. Positive culture by human rhabdomyosarcoma (RD) cell culture was selected for amplification and sequencing of the enterovirus VP1 region. Reference sequences of different genotypes were selected from Genbank, and a phylogenetic tree by neighbor joining (NJ) method was constructed using MAGA 11.0 software. MegaAlign in DNAStar software was used for VP1 sequence alignment and similarity analysis.Results:A total of 4 053 throat swab samples were collected from children with HFMD in Zibo city. The nucleic acid test results showed that the total positive rate of the samples was 70.91%, with the highest positive rate detected in children aged 3-6 years old in daycare (72.50%). The dominant serotype in 2014 was EV-A71, while the dominant serotypes in 2015, 2017, and 2018 were non-EV-A71 non-CVA16 enteroviruses. CVA16 was the dominant serotypes in 2016 and 2019. In the years after 2020, all dominant serotypes were non EV-A71 non-CVA16 enteroviruses. The incidence peaks of HFMD in the city occurred mainly from May to August with June as the peak. The differences in pathogen spectrum of HFMD in districts and counties were statistically significant ( χ2=48.841, P<0.05). The sequencing and comparative analysis of 63 positive isolates revealed that there were six types of enteroviruses in this city, i.e. CVA10 (28.57%), CVA16 (17.46%), CVA6 (7.94%), CV-A2 (6.35%), CV-A4 (12.70%) and EV-A71 (26.98%). The evolutionary analysis and similarity analysis showed that the EV-A71 strains of HFMD cases in Zibo city were highly concentrated and all belonged to C4a subgenotype. The nucleotide similarities of VP1 region among the strains ranged 92.50%-100.00%. All CVA16 strains of HFMD cases belonged to B1 subgenotype. B1a and B1b accounted for 36.36% and 63.64%, respectively. There was co-circulation of B1a and B1b and the nucleotide similarities in the VP1 region among the strains were 88.10%-100.00%. Conclusions:The pathogen spectrum of HFMD in Zibo city showed diversity from 2014 to 2022. From 2014 to 2019, the dominant strains of HFMD in Zibo city appeared alternately, gradually changed to non-EV-A71 non-CVA16 enteroviruses after 2020. Children aged 3-6 years old in childcare facilities was the key population. Testing of other enterovirus and VP1 typing should be carried out on focus to understand the local prevalence of HFMD and control the disease.