The invention provides an on-line calibration device and method for a leakage detection instrument. The leakage detection instrument (21) comprises a container (19) and a pressure meter (18). The calibration device is mainly composed of a gas supply system I (1), a gas supply system II (14), a needle valve (2), stop valves (3,6,8,10, 13,15 and 17), a pressure stabilizing chamber (5), a positive pressure standard leak (9), a calibration chamber (12), a pressure meter I (4), a pressure meter II (11), an air pumping system I (7) and an air pumping system II (16). The on-line calibration device can carry out calibration on the leakage detection instrument based on the constant volume pressure rise theory, is wide in calibration range and is used for calibrating various leak indication gases.