[Establishment of chemiluminescence immunoassay for detecting Staphylococcal enterotoxin B and C1].
OBJECTIVE
To establish chemiluminescence immunoassay (CLIA) for detecting staphylococcal enterotoxin B (SEB) and C1 (SEC1) and compare its sensitivity and stability with ELISA.


METHODS
The anti-SEB and SEC1 monoclonal antibodies (mAb) were purified by Q Sepharose Fast Flow chromatographic column. The alkaline phosphatase (AP) conjugated mAbs FMMU-SEB.D6 and FMMU-SEC1.C4 were used as detecting antibodies and the FMMU-SEB.B4 and FMMU-SEC1.G8 mAbs were used as coating antibodies in both methods. Phenolphthalein monophosphate (PMP) and lumigen APS-5 were employed as substrates for AP in ELISA and CLIA, respectively. The light was detected and measured by the GENios analyzer (TECAN Group Ltd.). The sensitivities and detect ranges of CLIA and ELISA methods were compared.


RESULTS
Compared with ELISA, CLIA was more sensitive (0.1 ng/mL vs 0.39 ng/mL) and timesaving. Furthermore, the liner range of CLIA was broader than that of ELISA (0.78-50 ng/mL vs 3.125-50 ng/mL).


CONCLUSIONS
CLIA for detecting SEB and SEC1 are established successfully which may be useful in food monitoring, epidemiology survey and detecting SE contaminated samples in environment.
Chen
Li-jie
LJ
Department of Immunology, Fourth Military Medical University, Xi'an 710032, China.
Liu
Zhen-shi
ZS
Dong
Bang-quan
BQ
Yang
Kun
K
Li
Qi
Q
Jin
Bo-quan
BQ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D004768:Enterotoxins / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D007118:Immunoassay / Q000379:methods*; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D012680:Sensitivity and Specificity; D013210:Staphylococcus; D013696:Temperature
D000911:Antibodies, Monoclonal; D004768:Enterotoxins; C031818:enterotoxin B, staphylococcal; C034959:enterotoxin C, staphylococcal