Antiphospholipid syndrome (APS) is an autoimmune disorder characterized by recurrent vascular thrombosis (VT) and/or pregnancy morbidity (PM) in the presence of persistent antiphospholipid antibodies (aPL), detected by lupus anticoagulant (LA), anticardiolipin (aCL) antibody, and/or anti-β₂ glycoprotein I (aβ₂GPI) antibody assays. These aPL, considered to be diagnostic markers and pathogenic drivers of APS, are a heterogeneous group of antibodies directed against anionic phospholipids, phospholipid-binding plasma proteins, and phospholipid-protein complexes. Although APS is currently considered as a single disease, it presents with a wide range of clinical symptoms and biological characteristics. The clinical diagnosis of APS in a patient with symptoms and signs is dependent upon the presence of a persistently positive result in an aPL assay. The tests recommended for detecting aPL are the standardized enzyme-linked immunosorbent assay (ELISA) to detect aCL and aβ₂GPI and clotting assays for LA performed according to the guidelines of the International Society on Thrombosis and Haemostasis. This chapter describes the standard laboratory test for the diagnosis of APS discussing the clinical and theoretical aspects of LA, aCL, and aβ₂GPI assays.