Highly sensitive microplate chemiluminescence enzyme immunoassay for the determination of staphylococcal enterotoxin B based on a pair of specific monoclonal antibodies and its application to various matrices.
A highly specific and sensitive microplate chemiluminescent enzyme immunoassay (CLEIA) was established and validated for the detection of staphylococcal enterotoxin B (SEB). A pair of monoclonal antibodies (mAbs) that recognizes different epitopes of SEB was selected from 20 SEB-specific mAbs, and the experimental conditions were examined and optimized for the development of the CLEIA. This method exhibited high performance with a dynamic range of 0.01-5 ng/mL, and the measured limit of detection (LOD) was 0.01 ng/mL. Intra- and interassay coefficient variations were all lower than 13% at three concentrations (0.2, 0.4, and 2 ng/mL). For specificity studies, when this method was applied to test staphylococcal enterotoxins A, C1, and D, no cross-reactivity was observed. It has been successfully applied to the analysis of SEB in a variety of environmental, biological and humoral matrices such as sewage, tap water, river water, roast beef, peanut butter, cured ham, 10% nonfat dry milk, milk, orange juice, and human urine and serum. The aim of this article is to show that the highly sensitive, specific, and simple microplate CLEIA, based on a pair of highly specific monoclonal antibodies, has potential applications for quantifying SEB in public health and military reconnaissance.
Liu
Fei
F
Department of Immunology, The Fourth Military Medical University, No. 17 Changle West Road, Xi'an 710032, Shaanxi Province, People's Republic of China.
Li
Yongming
Y
Song
Chaojun
C
Dong
Bangquan
B
Liu
Zhijia
Z
Zhang
Kui
K
Li
Haitao
H
Sun
Yuanjie
Y
Wei
Yuying
Y
Yang
Angang
A
Yang
Kun
K
Jin
Boquan
B
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002138:Calibration; D004768:Enterotoxins / Q000032:analysis* / Q000276:immunology; D005260:Female; D005504:Food Analysis; D006735:Horseradish Peroxidase / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D007700:Kinetics; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice
D000911:Antibodies, Monoclonal; D004768:Enterotoxins; C031818:enterotoxin B, staphylococcal; D006735:Horseradish Peroxidase