Muscle fibre conduction velocity is an important measurement in electrophysiology, both in the research laboratory and in clinical practice. It is usually measured by placing electrodes spaced at known distances and estimating the transit time of the action potential. The problem, common to all methods, is the estimation of this time delay. Several measurement procedures, in the time and frequency domains, have been proposed. Time-domain strategies usually require two acquisition channels, whereas some frequency-domain methods can be implemented using a single one. The method described operates in the time domain, making use of the autocorrelation function of the difference signal obtained from two needle electrodes and only one acquisition channel. Experimental results were obtained from the electromyogram of two biceps muscles (two adult male subjects, nine records each) under voluntary contraction, yielding an average of 3.58 m s−1 (SD=0.04 m s−1) and 3.37 m s−1 (SD=0.03 m s−1), respectively. Several tests showed that the proposed method works properly with electromyogram records as short as 0.3 s.