Open AccessDevelopment of a Clinical Chemiluminescent Immunoassay for Serum GPC3 and Simultaneous Measurements Alone With AFP and CK19 in Diagnosis of Hepatocellular Carcinoma
Author(s) -
Yu JuanPing,
Xu XiGuang,
Ma RuiJuan,
Qin ShiNi,
Wang CongRong,
Wang XiaoBo,
Li Ming,
Li MingSong,
Ma Qiang,
Xu WeiWen
Publication year - 2015
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.21733
Subject(s) - immunoassay , glypican 3 , chemiluminescent immunoassay , hepatocellular carcinoma , medicine , alpha fetoprotein , chemiluminescence , biomarker , antibody , coefficient of variation , antigen , likelihood ratios in diagnostic testing , cytokeratin , gastroenterology , receiver operating characteristic , immunology , chemistry , chromatography , immunohistochemistry , biochemistry
Background Glypican‐3 (GPC3) is an oncofetal antigen that shows great promise as a biomarker for diagnosis of hepatocellular carcinoma (HCC), but there is no reliable kit that can be used to detect it in clinics. The aim of this study is to develop a stable performance kit for GPC3 detection in clinics. Design and methods The paired antibodies were identified through cycle‐screening methods based on our previous research. Then, a double antibodies sandwich chemiluminescent immunoassay for detecting serum GPC3 was developed. The performance of the developed GPC3 diagnostic kit was evaluated by detecting the concentration of serum GPC3 and assessing its single or combined use with alpha fetoprotein (AFP) and cytokeratin 19 fragment (CK19) for HCC diagnosis. Results The assay demonstrated a linear range of 10–800 ng/ml, the cross‐reactivity rate at 0.018% (AFP), 0.020% (carcino‐embryonic antigen), and 0.021% (CK19), respectively. The minimum detectable concentration was 0.05 ng/ml; the intraassay coefficient of variation (CV) and interassay CV were both less than 10%, with good stability and reproducibility. GPC3 has a high sensitivity (54.2%) and specificity (99.4%) in diagnosing HCC. The level of GPC3 in HCC was robust higher than that in healthy or other liver diseases' sera (108.67 ± 230.04 ng/ml vs. 3.99 ± 7.68 ng/ml). The diagnostic sensitivity of GPC3 single or combined with CK19 and AFP for HCC was evaluated, and the rates were 54.2 and 90.6%, respectively. Conclusions An applicable chemiluminescent immunoassay with stable performance against GPC3 in diagnosing HCC has been established and the combination of GPC3 with CK19 and AFP could improve the diagnostic sensitivity for HCC.