The hole centers distance is difficult measured by using the conventional measurement method. In this paper,new detection method which combine a machine tool probe based on laser displacement sensor is proposed.With the machine tool coordinate system,the centers' relative coordinates are quickly and accurately gotten by using the approximation hole center location,and then the hole centers distance for the measured work piece can be also gotten. Comparing with the traditional measurement method,the present method is rapid,convenient,high precision,non-contact,on-line measurement. Finally,using a pair of standard ring gauge to verify the correctness of the method,checking measurement accuracy of the hole centers distance in the three coordinates measuring machine,measurement errors can reach micron level. The experiment prove the present method can efficiently applied to measure the hole center distance online.