Stigma and discrimination towards people living with HIV have been widely documented, and have extended their impact into the workplace. Stigmatising attitudes towards people living with HIV (PLHIV) in the workplace significantly hinder HIV prevention efforts and indirectly affect national development. This cross-sectional study was designed to determine the level of knowledge about HIV and AIDS and assess attitudes towards PLHIV among the general staff of Universiti Putra Malaysia (UPM), as well as to identify factors that are associated with it.