Objective：To analyze the cause of HBeAg （one of HBV markers） false-negative reaction during ELISA examination and to discuss its countermeasures. Methods： The patients who were HBsAg （＋） , HBcAb （＋） and HBeAg （-） by ELISA examination were further detected with the reagents of 4 different manufacturers in 2 steps： （1） The serum samples positive of HBsAg and HBcAb by ELISA using reagent A were further detected by qualitative analysis and Double-antibody sandwich ELISA using reagent A , B and C. （2） Reagent D （chemiluminescence method） was used to confirm the diagnosis if the results were positive or weakly positive of HBeAg, or the D values were within a specific range by ELISA using reagent A, B or C. The result of reagent D was taken as the final result. Results： The 274 sera negative of HBeAg were still negative when using reagent A, with the false-negative rate being 2.18%; 5 sera were positive when using reagent B, with false-negative rate being 0.36% ; and 6 sera （including the 5 positive ones using reagent B and 1 negative case using reagent A and B） were positive when using reagent C, without false-negative case. All the 6 positive samples were confirmed by chemiluminescence using reagent D. Conclusion： It is suggested that ELISA examination using reagent A or B can lead to false-negative results of HBeAg. HBsAg （＋）, HBcAb （＋） and HBeAg （-） sera in ELISA examination should be examined with combinations of reagents of different manufacturers.