Laser induced fluorescence (LIF) technique has been demonstrated as a powerful technology for analyzing the contamination of petroleum due to its excellent attributes of rapid analysis speed and slight sample preparation. This study focuses on the monitoring application of LIF in petroleum hydrocarbon-contaminated soils by establishing the three-dimensional diffusion models. In this paper, to improve the analysis accuracy, the effects of soil matrix difference for fluorescence intensities were considered. In order to validate the practicability of LIF, the longitudinal penetration laws and the lateral diffusion laws of diesel oil in different humidity soils were analyzed. These laws indicate that the longitudinal penetration depth decreases and the lateral diffusion range increases with the increase of soil moisture. Then, the three-dimensional diffusion models were established, the relative standard deviation (RSD) of the predictions for diesel oil in different soil moisture are 5.09%, 9.62%, 7.92%, and the contaminated volumes of soils by diesel oil are 233.90 cm3, 332.70 cm3, and 660.05 cm3, respectively. These results express that the soil moisture extends the extent of diesel-contaminated soils. The present work shows the feasibility of LIF technique for the field monitoring of petroleum.