Heart rate is quite regular during sinus (normal) rhythm (SR) originating from the sinus node. In contrast, heart rate is usually irregular during atrial fibrillation (AF). Complete atrioventricular block with an escape rhythm, ventricular pacing, or ventricular tachycardia are the most common exceptions when heart rate may be regular in AF. Heart rate variability (HRV) is the variation in the duration of consecutive cardiac cycles (RR intervals). We investigated the utility of HRV parameters for automated detection of AF with machine learning (ML) classifiers. The minimum redundancy maximum relevance (MRMR) algorithm, one of the most effective algorithms for feature selection, helped select the HRV parameters (including five original), best suited for distinguishing AF from SR in a database of over 53,000 60 s separate electrocardiogram (ECG) segments cut from longer (up to 24 h) ECG recordings. HRV parameters entered the ML-based classifiers as features. Seven different, commonly used classifiers were trained with one to six HRV-based features with the highest scores resulting from the MRMR algorithm and tested using the 5-fold cross-validation and blindfold validation. The best ML classifier in the blindfold validation achieved an accuracy of 97.2% and diagnostic odds ratio of 1566. From all studied HRV features, the top three HRV parameters distinguishing AF from SR were: the percentage of successive RR intervals differing by at least 50 ms (pRR50), the ratio of standard deviations of points along and across the identity line of the Poincare plots, respectively (SD2/SD1), and coefficient of variation—standard deviation of RR intervals divided by their mean duration (CV). The proposed methodology and the presented results of the selection of HRV parameters have the potential to develop practical solutions and devices for automatic AF detection with minimal sets of simple HRV parameters. Using straightforward ML classifiers and the extremely small sets of simple HRV features, always with pRR50 included, the differentiation of AF from sinus rhythms in the 60 s ECGs is very effective.