To analyze the methodology of serum alpha fetoprotein (AFP) assay by high-throughput enzyme-linked immunosorbent assay (ELISA) and appraise its clinical value.

A total of 170 serum samples of patients (hepatic carcinoma, hepatitis and hepatic cirrhosis patients) and 200 serum samples of healthy people were recruited from Qingdao Central Hospital from March to December 2013 in this study. The linear range, precision, accuracy, reference interval and diagnostic performance for primary liver cancer were estimated in accordance with EP5-A2 files and EP15-A2 files from the American association of Clinical Laboratory Standards Institute and related literatures. Compared with the electrochemical luminescence assay was performed to evaluated the detection accuracy and clinical value of high-throughput ELISA.

The linear range of high-throughput ELISA for AFP was 0.0-200.0μg/L. The intra-batch coefficient of variation (CV ) of different concentrations (low, medium and high) were 4.9%, 3.1%, 3.7% respectively, the inter-batch CVs of different concentrations were 3.8%, 1.5%, 2.1% respectively, while total CVs of different concentrations were 5.0%, 6.0%, 5.0% respectively. Compared with the results of the electrochemical luminescence, the relative bias was 7.9%. The recovery rate was 91%-109%, 97.76% on average. The reference interval of serum AFP detected by high-throughput ELISA in our laboratory was 0.00~7.20 μg/L. The diagnostic threshold of primary liver cancer was 195.7μg/L, with sensitivity of 39.5% and specificity of 93.3%.

This study established standard operating procedures for the quantitative detection of serum AFP by a high-throughput ELISA method. High-throughput ELISA assay for quantitation of serum AFP is an accurate, stable performance in diagnosis of primary liver cancer.