The long-term management of chronic hepatitis B (CHB) involves multiple steps, including screening, diagnosis, disease staging, monitoring of antiviral therapy, assessment of treatment discontinuation and risk of relapse, and surveillance for cirrhosis and hepatocellular carcinoma. Laboratory biomarkers are central to clinical evaluation and decision-making in CHB. With advances in hepatitis B virus (HBV) research and diagnostic technologies, laboratory biomarkers for CHB have evolved from conventional serologic and virologic markers to newer markers that reflect viral transcriptional activity, antigen expression characteristics, and the status of the viral reservoir. While improving diagnosis and treatment rates remains a fundamental goal in CHB care, the recognition of functional cure as an important therapeutic endpoint has further increased the need for biomarkers that can assess residual viral activity, stratify relapse risk after treatment withdrawal, and guide long-term follow-up strategies. In this review, we summarize the evidence base and clinical application characteristics of conventional and novel laboratory biomarkers in CHB, and further discuss the roles of reflex testing and laboratory test utilization management in optimizing clinical pathways for CHB. Overall, the optimization of laboratory testing in CHB should not depend on simply increasing the number of assays, but rather on the rational integration of biomarkers according to specific clinical questions, so as to improve diagnostic and management efficiency, reduce unnecessary testing, and better support stratified management, long-term follow-up, and the assessment of functional cure.