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, mono-clonal antibodies (McAbs) against pepsinogen...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, mono-clonal antibodies (McAbs) against pepsinogen Ⅰ (PG Ⅰ) and PG Ⅱ were co-coated in 96 microtitration wells, and tracer McAbs against PG Ⅰ 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 μg/L for PG Ⅰ and 0.05 μg/L for PG Ⅱ. The assay range was 5.0-320.0 μg/Lfor PG Ⅰ and 1.0-55.0 μg/L for PG Ⅱ. The average re-covery rate was 102.7% for PG Ⅰ and 98.8% for PG Ⅱ. Sera from healthy controls and patients with gastric dis-ease were analyzed. The PG detected by dual-label as-say 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.展开更多
基金Supported by The Program of Social Development Fund from Jiangsu Science and Technology Department, No. BS2006015the Program of Health Department of Jiangsu Province, No. H200856
文摘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, mono-clonal antibodies (McAbs) against pepsinogen Ⅰ (PG Ⅰ) and PG Ⅱ were co-coated in 96 microtitration wells, and tracer McAbs against PG Ⅰ 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 μg/L for PG Ⅰ and 0.05 μg/L for PG Ⅱ. The assay range was 5.0-320.0 μg/Lfor PG Ⅰ and 1.0-55.0 μg/L for PG Ⅱ. The average re-covery rate was 102.7% for PG Ⅰ and 98.8% for PG Ⅱ. Sera from healthy controls and patients with gastric dis-ease were analyzed. The PG detected by dual-label as-say 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.
