Abstract: laboratory simulation test was used for this research, the relation between population density of EPN in soil and the individual number in detection tube was studied. Collection time and the distance between detection tubes were studied by one way test. The results showed that collection time and the distance between detection tubes affected the individual number in detection tube significantly. 48 hours was the best Collection time and 8 cm was the best distance between detection tubes for the EPN collection. The result showed that the individual number in detection tube was related to the population density of EPN in soil, the highest R² was 0.9364. This research provide a new method for qualitative and quantitative of EPN detection.