PIVKA-II is well known as a tumor maker of hepatocellular carcinoma (HCC). We measured serum PIVKA-II concentrations with a commercially available PIVKA-II immunoassay kit (Picolumi PIVKA-II: Eisai Co., Ltd., Tokyo) using Electrochemiluminescence Immunoassay (ECLIA). ECLIA system is a novel immunoassay system using a Ruthenium (II) Tris (bipyridyl) luminesced by electric energy. Cut off value was 40 mAU/ml by receiver-operating characteristic curves as a tumor maker for HCC. Eighty-nine out of 142 (62.2%) patients with HCC had elevated serum PIVKA-II levels and seventeen out of 36 (47.2%) patients whose tumor size was below 2 cm in diameter showed high PIVKA-II levels. We determined the serial changes in serum PIVKA-II levels of two patients with HCC following initial therapy. In these patients, elevations of serum PIVKA-II levels determined by ECLIA system preceded the HCC relapse detected by imaging diagnostic procedures. In summary, this assay system is suitable for detecting small increases in PIVKA-II concentrations. Determination of PIVKA-II by this assay system is found to be useful for the early detection of HCC.