Most of the research of mental fatigue evaluation mainly concentrated on some indexes that require sophisticate and large instruments which make the detection of mental fatigue cumbersome, time-consuming, and difficult to apply on a large scale. A quick and sensitive mental fatigue detection index is necessary so that mental workers can be alerted in time and take corresponding countermeasures. But to date, no studies have compared the sensitivity of common objective evaluation indexes. To solve these problems this study recruited 56 human subjects. These subjects were evaluated using six fatigue indexes: the Stanford sleepiness scale, digital span, digital decoding, short-term memory, critical flicker fusion frequency (CFF), and speed perception deviation. The results of fatigue tests before and after mental fatigue were compared, and a one-way analysis of variance (ANOVA) was performed on the speed perception deviation. The result indicated the significance of this index. Considering individual differences, the relative fatigue index (RFI) was proposed to compare the sensitivity of the indexes. The results showed that when the self-rated fatigue grade changed from non-fatigue to mild fatigue, the ranges of RFI values for digital span, digital decoding, short-term memory and CFF were 0.175–0.258, 0.194–0.316, 0.068–0.139, and 0.055–0.075, respectively. Correspondingly, when the self-rated fatigue grade changed from non-fatigue to severe fatigue, the ranges of RFI values for the above indexes were 0.175–0.258, 0.194–0.316, 0.068–0.139, and 0.055–0.075, respectively. These results suggest that the sensitivity of the digital decoding, digital span, short-term memory, and CFF decreased sequentially when the self-evaluated fatigue grade changed from no fatigue to mild or severe fatigue. The RFI individuality of the speed perception deviation is highly variable and is not suitable as an evaluation index. In mental fatigue testing, digital decoding testing can provide faster, more convenient, and more accurate results.