Objective: To analyze the relationship between Toll like receptor ( TLR ) 9 and TLR 7 gene polymorphisms among children with hand-foot-mouth disease in a medical institution of Jinan City from 2020 to 2023, so as to provide reference for prevention of hand-foot-mouth disease. Methods: Data of 284 preschool children aged 3-6 with hand-foot-mouth disease, including the season pattern, clinical manifestations, pathogen distribution, were collected in department of pediatric infectious diseases of the Second Children & Women s Healthcare of Jinan from 2020 to 2023. The distribution conditions of TLR 9 and TLR 7 genotypes of enterovirus 71 (EV71) subtype and coxsackievirus A16 (CA16) subtype were compared by Chi square test. Hardy Weinberg equilibrium test was used to characteristic sample population distribution, and Logistic regression analysis in the additive genetic model was adopted to evaluate the relationship between TLR 9 and TLR 7 gene polymorphisms with hand-foot-mouth disease subtype. Results: The time to admission after symptom onset with hand-foot-mouth disease among 284 children was mostly >2-4 d ( 45.42% ), followed by ≤2 d ( 44.72% ). The season pattern was mostly in summer (58.10%). The severity was mostly mild ( 69.72% ), and the main clinical manifestations were skin rash (100.00%) and fever (65.49%). The majority pathogen was EV71 (60.56%). Hardy Weinberg equilibrium test showed good population representation ( P >0.05). Logistic regression analysis was applied to analyze the relationship between TLR9 rs 187084 locus and hand-foot-mouth disease subtypes under the additive genetic model after adjusting for age and gender of the study population, and it was found that compared with those without C allele, each additional C allele increased the risk of CA16 subtype hand-foot-mouth disease by 109% ( OR=2.09, 95%CI =1.13-3.27). Analysis of relationship between TLR 7 rs 3853839 locus and hand-foot-mouth disease subtype showed that compared with those without C allele, the risk of EV71 subtype hand-foot-mouth disease increased by 57% for each additional C allele ( OR=1.57, 95%CI = 1.10 -2.11) ( P <0.05). Conclusions From 2020 to 2023, children with hand-foot-mouth disease in a medical institution of Jinan City is mostly mild and caused by EV71 infection. The polymorphism of rs 3853839 locus of TLR 7 may be related to EV71 infection in children with hand-foot-mouth disease, and the polymorphism of rs 187084 locus of TLR 9 may be related to CA16 infection.