Simultaneous detection of different serum pepsinogens and its primary application

doi:10.3748/wjg.v16.i24.3072

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Jun 28, 2010 (publication date) through Nov 4, 2024

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World Journal of Gastroenterology

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1007-9327

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Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

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World J Gastroenterol. Jun 28, 2010; 16(24): 3072-3077
Published online Jun 28, 2010. doi: 10.3748/wjg.v16.i24.3072

Simultaneous detection of different serum pepsinogens and its primary application

Jue Zhang, Ji-Zhong Guo, Hua-Long Xiao, Lan Zhu, Hai-Yan Liu, Yi Zhang, Biao Huang

Jue Zhang, Lan Zhu, Hai-Yan Liu, Yi Zhang, Biao Huang, Key Laboratory of Nuclear Medicine, Ministry of Public Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Wuxi 214063, Jiangsu Province, China

Ji-Zhong Guo, Hua-Long Xiao, Wuxi People’s Hospital, Wuxi 214023, Jiangsu Province, China

Author contributions: Zhang J, Huang B and Guo JZ designed the study; Zhang J interpreted the data and wrote the manuscript; Zhu L, Liu HY and Zhang Y performed the majority of experiments and data analysis; Xiao HL collected the samples.

Supported by The Program of Social Development Fund from Jiangsu Science and Technology Department, No. BS2006015; the Program of Health Department of Jiangsu Province, No. H200856

Correspondence to: Biao Huang, PhD, Professor, Key Laboratory of Nuclear Medicine, Ministry of Public Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Wuxi 214063, Jiangsu Province, China. jyswjs@yahoo.com.cn

Telephone: +86-510-85520770 Fax: +86-510-85520770

Received: February 17, 2010
Revised: March 15, 2010
Accepted: March 22, 2010
Published online: June 28, 2010

Abstract

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 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/L for PG I and 1.0-55.0 μg/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.

Keywords: Serum pepsinogen; Simultaneous detection; Time-resolved fluoroimmunoassay