The objective of this study was to investigate the association between physical fitness and body mass index categories (obesity, OB; overweight, OW; normal-weight, NW; and underweight, UW) in prepubertal children. Anthropometric and physical fitness characteristics were collected from a convenience sample of 30472 Italian schoolchildren (6-11 years old). Six field-based tests were used: Lé ger, agility shuttle, long jump, frontal throw of the basketball, Sit & Reach and standing balance. Significant differences were found in the anthropometric characteristics, physical fitness and weight status prevalence between girls and boys (p<0.05) and, except for flexibility, by age class (p<0.05). Obese children performed worse than their NW counterparts in aerobic capacity (p<0.001), agility (p<0.001), muscular power of the lower limb (p<0.001) and balance (p<0.001). Conversely, children with obesity showed greater upper limb power than NW children (p<0.001). The discrepancy in physical fitness between OB and NW children increased in older girls (flexibility, p = 0.002; muscular power of the lower and upper limb, p = 0.002 and p = 0.005) and boys (aerobic capacity, p = 0.009; agility, p = 0.006; standing balance, p = 0.019; muscular power of the lower and upper limb, p<0.001 and p = 0.011) compared to their younger counterparts. On the other hand, UW children performed worse than NW children mainly in terms of muscular power of the arms (p<0.001). Additionally, there was an increasing disparity in the frontal throw test scores of UW and NW girls (p = 0.003) and boys (p = 0.011) in older children compared to younger children. In conclusion, the effect of body mass index on children's physical fitness intensifies with age. OB and OW negatively affect aerobic capacity, agility, lower limb power and balance but positively affect upper limb power. UW negatively affects upper limb power. This study underscores the importance of preventing childhood OW, OB, and UW in early life to promote children's health and proper fitness development.