The invention provides a measurement and regulation method for a rough rolling working roll gap deviation. The measurement and regulation method comprises the following steps of: after the rolls of a first rough mill are changed, putting two copper bars of which the diameter is Phi 18-22mm in parallel with a rolling line into a roll gap; manually pressing down until the copper bars can not be pressed; respectively measuring the thickness of each laminated copper bar by a micrometer; if the thickness difference of the two copper bars is less than or equal to 0.3mm, carrying out horizontal roller calibration; if the thickness difference of the two copper bars is more than 0.3mm, calibrating again; selecting two copper bars of which the diameter is Phi 8-12mm, and carrying out secondary calibration according to the above steps; charging one short ruler board blank into a heating furnace for real rolling verification; carrying out rough rolling and pressing for 10-15mm; observing the practical rolling situation of a slab; remelting and rolling the calibrated slab; and according to the rolled material shape, regulating the horizontal roll gap of the rough rolling working roll. According to the measurement and regulation method disclosed by the invention, the practical deviation of the roll gap of the measurement side of the rough mill working roll can be accurately measured, the roll gap calibration and presetting precision of the roll gap can be improved, the middle blank shape is guaranteed to be rolled to be flat, and the steel clamping accidents caused by middle blank lateral bending are avoided.