Abstract: Background and Objectives: Chemically synthesized surfactants used as positive controls in assessing for biosurfactant production are expensive and not readily available in microbiology laboratories in Nigeria. The objectives of this study included comparing the response values of selected detergents to screening methods used in assessing for biosurfactant production with those of standard surfactants. Materials and Methods: Glycerol-mineral salts medium was used for culturing Pseudomonas fluorescens for biosurfactant production. At the end of the production period, the medium and 1% of selected detergents (Klin, Ariel and Cussons Morning fresh) were screened for surfactant activity using surface tension, foaming capacity, oil spread diameter and drop collapse activity. Results: The surface tension of the medium used for culturing P. fluorescens for biosurfactant production was reduced from 55.21-34.63 mN m–1, the foaming capacity was 58.83% and the oil spread diameter was about 15 mm. The detergent solution having the least surface tension and the largest oil spread diameter was cussons morning fresh (17.84 mN m–1 and 70 mm, respectively), while the detergent having the highest foaming capacity was Ariel (82.6%). The drop collapse activity of the medium and the detergent solutions were all positive. Conclusion: The results obtained for the detergents were comparable with those of some chemically synthesized surfactants. This indicates that readily available and inexpensive detergents can be used in place of chemically synthesized surfactants as positive controls in assessing for biosurfactant production.