Acquired Immunodeficiency Syndrome (AIDS) is a severe infectious disease induced by human immuno deficiency virus (HIV). Laboratory testing plays an important role in the diagnosis of HIV/AIDS. In general, laboratory testing includes detection of virus antibodies and antigens, virus RNA, immune cells (CD4+, CD8+) and anti-HIV drug resistance. During the past twenty years, great progress has been made in laboratory testing. As research on HIV/AIDS has advanced and biotechnology has developed rapidly, different methods of testing have been discovered. In recent years, the application of molecular biotechnology and immunology has led to important advances for epidemiological surveys, clinical diagnosis and treatment of HIV. The existence of a testing method with high sensitivity and specificity is not only helpful for early diagnosis and prediction, monitoring and evaluation of therapeutic efficacy, but can also reduce the risk of false-negative results. HIV laboratory testing is now developing towards a simple, rapid, sensitive, accurate and automatic way of diagnosing this condition.