Objective To study on the improvement of incubation time and plate load capacity for molds enumeration. Methods Mold quality control samples were tested according to GB 4789.15—2016 National food safety standard-Food microbiological examination-Enumeration of moulds and yeasts, with additional colony counting performed on the second day of incubation. The plate size was increased from 90 mm to 150 mm, and statistical analysis was employed to assess the differences between colony counts and the reference value of the quality control samples. Results Significant discrepancies were observed between the fifth-day counting result at the critical dilution (1:10) and the reference value. A sharp decline in colony counts was noted when the colony-forming units exceeded 30 CFU per 90 mm plate, which was defined in this study as the maximum load capacity of mold plates under acceptable accuracy. By adopting 150 mm plates with a 2-day incubation period, the original load capacity was overcome, yielding counting results that aligned well with the reference value. Under these optimized conditions, the linear coefficient of determination (R²) between sample means and dilution factors improved from R²₉₀=0.614 to R²₁₅₀=0.998. Conclusion The optimized testing conditions effectively addressed the issues of colony overgrowth and poor linearity in mold plate count experiments, thereby enhancing the accuracy of test results.