We propose a new method - twitch force - for estimation of the muscle behavior during voluntary contraction for assessing localized muscle fatigue. The proposed method uses the sEMG peaks as input and the measured force as output. The twitch force, which is a transfer function to generate force, was estimated during fatiguing contraction. We verified the estimated twitch force based on the measured results with electrical stimulation. The participants performed isometric little finger flexion until exhaustion. SEMG was recorded on the flexor digiti minimi brevis muscle for the proposed method and the electrical stimulation electrodes on the ulnar nerve induced involuntary contraction for reference. As the muscle fatigue level increased, the twitch peaks decreased in both methods. The proposed method can be widely used in the quantitative analysis of muscle fatigue during voluntary contraction.