[Purpose] Intelligent emotion assessment systems have been highly successful in a variety of applications, such as e-learning, psychology, and psycho-physiology. This study aimed to assess five different human emotions (happiness, disgust, fear, sadness, and neutral) using heart rate variability (HRV) signals derived from an electrocardiogram (ECG). [Subjects] Twenty healthy university students (10 males and 10 females) with a mean age of 23 years participated in this experiment. [Methods] All five emotions were induced by audio-visual stimuli (video clips). ECG signals were acquired using 3 electrodes and were preprocessed using a Butterworth 3rd order filter to remove noise and baseline wander. The Pan-Tompkins algorithm was used to derive the HRV signals from ECG. Discrete wavelet transform (DWT) was used to extract statistical features from the HRV signals using four wavelet functions: Daubechies6 (db6), Daubechies7 (db7), Symmlet8 (sym8), and Coiflet5 (coif5). The k-nearest neighbor (KNN) and linear discriminant analysis (LDA) were used to map the statistical features into corresponding emotions. [Results] KNN provided the maximum average emotion classification rate compared to LDA for five emotions (sadness - 50.28%; happiness - 79.03%; fear - 77.78%; disgust - 88.69%; and neutral - 78.34%). [Conclusion] The results of this study indicate that HRV may be a reliable indicator of changes in the emotional state of subjects and provides an approach to the development of a real-time emotion assessment system with a higher reliability than other systems.