Objective To observe the ability of inactivating human immunodeficiency virus type 1 (HIV-1) in whole blood by visible light combination with methylene blue (MB) at different concentrations. Methods HIV-1 was used as the test virus. The contaminated blood was treated by MB, visible light (640 nm), combined with MB and visible light (640 nm). The test of MT4 cell infection was used to evaluate the virus inactivation efficacy. Results After being treated by MB at the concentrations of 5, 10, and 15 ?mol/L, and then irradiated respectively by visible light (40 000 Lux, 640 nm) for 30, 20, and 10 min, all the added indicated virus at the titer of 10 5.78 TCID 50 of HIV-1 could be inactivated absolutely. Conclusion Methylene blue photochemical method can inactivate HIV-1 in blood effectively.