Drug addiction is a chronic disease that causes serious physical and psychological harm to the individual, as well as damage to society. Biomarkers are indicators that differ significantly between healthy people and patients and are important for the diagnosis and treatment of diseases. The increasing incidence of drug addiction in recent years and the wider variety and number of new drugs has led to an increase in the study of biomarkers of drug addiction and their analysis in different biological samples. This paper reviews biomarkers of drug addiction in hair,blood, urine, and feces, to help to explore the biological signatures of drug addiction and facilitate its clinical diagnosis and treatment.