Simultaneous detection of different serum pepsinogens and its primary application

AIM: To develop the simple, rapid and sensitive dual-label time-resolved fluoroimmunoassay for pepsinogens in human serum. METHODS: Based on two-site sandwich protocol, monoclonal antibodies (McAbs) against pepsinogen I (PG I) and PG II were co-coated in 96 microtitration wells, and tracer McAbs against PG 1 and PG II were labeled with europium (Eu) and samarium (Sm) chelate, respectively. Diluted serum samples of Eu3+- and Sm3+-McAbs were added into microtitration wells simultaneously. After washing, fluorescence of bound Sm3+ and Eu3+ tracers was detected. RESULTS: The detection limit was 0.2 mu g/L for PG I and 0.05 mu g/L for PG II. The assay range was 5.0-320.0 mu g/L for PG I and 1.0-55.0 mu g/L for PG II. The average recovery rate was 102.7% for PG I and 98.8% for PG II. Sera from healthy controls and patients with gastric disease were analyzed. The PG detected by dual-label assay was in good agreement with that detected by single-label assay or by enzyme-linked immunosorbent assay. CONCLUSION: Dual-label assay can provide high-throughput serological screening for gastric diseases. (C) 2010 Baishideng. All rights reserved.