Simultaneous detection of different serum pepsinogens and its primary application.
OBJECTIVE
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 Eu(3+)- and Sm(3+)-McAbs were added into microtitration wells simultaneously. After washing, fluorescence of bound Sm(3+) and Eu(3+) tracers was detected.


RESULTS
The detection limit was 0.2 microg/L for PG I and 0.05 microg/L for PG II. The assay range was 5.0-320.0 microg/L for PG I and 1.0-55.0 microg/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.


CONCLUSIONS
Dual-label assay can provide high-throughput serological screening for gastric diseases.
Zhang
Jue
J
Key Laboratory of Nuclear Medicine, Ministry of Public Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Wuxi 214063, Jiangsu Province, China.
Guo
Ji-Zhong
JZ
Xiao
Hua-Long
HL
Zhu
Lan
L
Liu
Hai-Yan
HY
Zhang
Yi
Y
Huang
Biao
B
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005063:Europium / Q000737:chemistry; D015200:Fluoroimmunoassay / Q000379:methods* / Q000592:standards; D005767:Gastrointestinal Diseases / Q000097:blood / Q000175:diagnosis; D006801:Humans; D057230:Limit of Detection; D010435:Pepsinogens / Q000097:blood*; D012493:Samarium / Q000737:chemistry; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D010435:Pepsinogens; D012493:Samarium; D005063:Europium