Abstract: ［Abstract］Objective　To compare and discuss the different results among four kinds of experimental methods（saline water method，Antiglobulin method，Polybrene method ，Microcolumn gel test） in Rh(D) negative confirmation experiment， and analyze their effects . Methods　We selected 96 cases whose results of Rh(D) blood typing in the early screening specimen were negative in the department of blood transfusion of the first people's hospital of yunnan province between January 2012 and December 2012. And we used saline wator method,antiglobulin method，polybrene method， microcolumn gel test to make Rh(D) the negative confirmation experiment and typing experiments at the same time. Results 96 cases were all confirmed the negative results through Saline method. 84 cases of 96 were confirmed the negative results through Polybrene method. 82 cases of 96 were confirmed the negative results through Anti-human globulin test. 81 cases of 96 were confirmed the negative results through Microcolumn gel test. Conclusions　Saline water method is simple but has poor specificity．Polybrene method is simple and rapid， but it will miss the IgG antibody of weaker antibody titer. The operation of antiglobulin method is complex， but its sensitivity and specificity are good. Microcolumn gel test for detection of IgG antibody in the serum has the characteristics of high sensitivity，strong specificity，and simple operation . In practice， combination of various methods can more guarantee the accuracy of the results.