Research was conducted to assess the practical use of an ATP bioluminescence assay to evaluate the bacteriological quality of raw milk. Filtration was used to precondition samples before ATP determination, which was measured in relative light units (RLUs). The Lumac ATP bioluminescence assay results were compared with standard plate counts (SPCs) of samples to estimate the microbial load for 246 raw milk samples that were split and either tested immediately or subjected to two preliminary incubation temperatures, 12.8 and 15.6 degrees C, for 18 h. Linear regression analysis procedures were used to analyze the data. Preincubation treatments were analyzed separately. For all treatments, linear regression coefficients were significantly different from zero (P < 0.01). The R2 values calculated using log10-transformed SPC and log10-transformed RLUs for samples tested immediately and samples preliminarily incubated at 12.8 and 15.6 degrees C were 0.58, 0.78, and 0.80, respectively. The R2 for all samples combined was 0.78. Differences in regressions among treatments were tested using a multiple slope and intercept model. Treatment intercepts and slopes were significantly different (P < 0.01). A linear regression equation was used to predict SPC from ATP values. Comparison of predicted values with actual SPCs indicated that ATP could be useful in predicting SPC in raw milk.