This paper presents a camera-based prototype sensor for detecting fatigue and drowsiness in drivers, which are common causes of road accidents. The evaluation of the detector operation involved eight professional truck drivers, who drove the truck simulator twice—i.e., when they were rested and drowsy. The Fatigue Symptoms Scales (FSS) questionnaire was used to assess subjectively perceived levels of fatigue, whereas the percentage of eye closure time (PERCLOS), eye closure duration (ECD), and frequency of eye closure (FEC) were selected as eye closure-associated fatigue indicators, determined from the images of drivers’ faces captured by the sensor. Three alternative models for subjective fatigue were used to analyse the relationship between the raw score of the FSS questionnaire, and the eye closure-associated indicators were estimated. The results revealed that, in relation to the subjective assessment of fatigue, PERCLOS is a significant predictor of the changes observed in individual subjects during the performance of tasks, while ECD reflects the individual differences in subjective fatigue occurred both between drivers and in individual drivers between the ‘rested’ and ‘drowsy’ experimental conditions well. No relationship between the FEC index and the FSS state scale was found.