Purpose <p>Early detection of cardiac amyloidosis (CA) remains challenging, and a PET tracer capable of directly visualizing amyloid deposits could improve diagnosis. Given the shared <i>β</i>-sheet structure of amyloid deposits in CA and Alzheimer’s disease (AD), this study aimed to evaluate the suitability of the AD amyloid tracer [¹⁸F]Florbetazine for non-invasive detection of CA via preclinical assessments and preliminary clinical evaluation.</p> Methods <p>Specific binding and affinity of [¹⁸F]Florbetazine to amyloid light-chain (AL) and transthyretin (ATTR) deposits were assessed via in vitro autoradiography on human myocardial sections. Pharmacokinetics and biodistribution were characterized by dynamic micro-PET/CT imaging in healthy Sprague Dawley (SD) rats. Whole-body PET/CT imaging was performed in two healthy volunteers to estimate normal-organ background, and in a single patient with biopsy-proven AL amyloidosis to preliminarily explore diagnostic feasibility.</p> Results <p>Autoradiography confirmed specific binding of [¹⁸F]Florbetazine to both AL and ATTR deposits with high binding affinities (<i>K</i>_d = 20.85 nM and 54.10 nM, respectively). In rats, [¹⁸F]Florbetazine exhibited rapid myocardial and renal uptake followed by efficient clearance from normal tissues. In healthy volunteers, background myocardium and renal activity was minimal by 40&#xa0;min post-injection. In the AL amyloidosis patient, delayed-phase PET/CT imaging (60–70&#xa0;min) revealed intense myocardium and renal uptake, correlating with renal biopsy findings.</p> Conclusions <p>These integrated preclinical and preliminary clinical evaluation demonstrates that [¹⁸F]Florbetazine binds cardiac amyloid with high affinity and displays favorable pharmacokinetics, warranting further investigation in larger, systematically designed clinical studies.</p> Trial registration <p>Clinical trial registry NCT06593626 (registered 24 May 2024) and NCT06725706 (registered 5 Dec 2024).</p>