Objective of the present study is to determine the knowledge, attitude, and practice of universal precautions amongst medical laboratory workers in the two Colleges of Medicine and their Teaching hospitals in Lagos State, Nigeria. Cross-sectional study of health care workers was Conducted using a standardized self-administered questionnaire, which enquired about knowledge, attitude and practices of universal precautions. The hepatitis B vaccination statuses were also determined. 300 questionnaires were administered and 154 of them were returned giving a response rate of 51.3%. Participants included; Medical laboratory scientist 55.8%(n=86), Laboratory at-tendants 23.3%(n=36), Medical laboratory technician 10.4%(n=16), Post-graduate students 7.8%(n=12) and Medical doctors 2.59%(n=4). All the participants wear gloves during laboratory work but 81.2% wear a single pair. Ny-lon gloves were commonly used (57%) followed by latex gloves (43%). 17.5 % of the partici-pants claimed to know what to do if exposed to infection. (i) 45.6% of the participants eat in the laboratory, (ii) 47.0% of them store foods and water in the refrigerators meant for stor-age of body fluids and chemicals, (iii) 31.5% of them put on cosmetics in the laboratory, (iv) 12.6% smoke and sniff in the laboratory, (v) 10.0% cut their finger nails with teeth and put their biros in their mouths in the laboratory. (vi) 36.5% do not know that tissues fixed in formalin can transmit infections. (vii) 91.5% are not immunized against hepatitis B virus (HBV). (viii) 99.0% of them do not take shower immediately after laboratory work. (ix) 82.0% of the participants do not feel that the use of masks is necessary in laboratory. It is concluded that the knowledge, attitude, perception, and compliance with universal pre-cautions amongst these highly exposed laboratory workers are poor.