In this study, an artificial autoassociative neural network (AANN) was used online to detect deviations from normal antibiotic production fermentation using conventional process variables. To improve the efficiency of extracting hidden information contained in multidimensional process variables, and to finally render the AANN adequate for fault detection, we explored the following methods: selection of process variables; preprocessing of data that involved normalizing the training data of the AANN; and evaluation of data that involved assessing the output of the AANN. A method for fault detection in virginiamycin M and S production by Streptomyces virginiae was successfully developed based on these techniques.