Monoclonal-based enzyme-linked immunosorbent assay for the detection of zearalenone in cereals.
A monoclonal antibody against zearalenone (ZEA) was produced and used successfully to develop a direct competitive enzyme-linked immunosorbent assay (DC-ELISA) for the analysis of ZEA in cereals. This DC-ELISA had a limit of detection of 0.15 +/- 0.02 microg l(-1) and an IC50 value of 1.13 +/- 0.16 microg l(-1). Matrix interference was minimized by dilution of the sample extract before ELISA assays. Aqueous methanol (80%) gave good extraction efficiencies, and the recovery from spiked rice, barley, and corn samples averaged between 87 and 112%. Although ZEA was detected in seven (9%) of 80 rice samples and in eight (16%) of 50 barley samples, the concentration of ZEA in samples was around or below the limit of detection of DC-ELISA. Among 38 corn samples, ZEA was detected in nine (24%) samples in the range 41.0-909.8 microg kg(-1). Re-analysis of the ELISA-positive corn samples by high-performance liquid chromatography (HPLC) confirmed that seven (18%) corn samples were positive. The ZEA results for corn showed very good agreement between DC-ELISA and a commercial AgraQant zearalenone kit (r2 = 0.98). Thus, the monoclonal antibody-based DC-ELISA could be applied to the preliminary screening of ZEA contamination when analysis of a large sample number is needed.
Thongrussamee
T
T
Division of Applied Life Science, Brain Korea 21 Program, Graduate School of Gyeongsang National University, Jinju, Gyeongnam, Korea.
Kuzmina
N S
NS
Shim
W-B
WB
Jiratpong
T
T
Eremin
S A
SA
Intrasook
J
J
Chung
D-H
DH
D000911:Antibodies, Monoclonal / Q000302:isolation & purification / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002523:Edible Grain / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D004968:Estrogens, Non-Steroidal / Q000032:analysis* / Q000276:immunology; D005506:Food Contamination / Q000032:analysis*; D001467:Hordeum / Q000737:chemistry; D006801:Humans; D012275:Oryza / Q000737:chemistry; D015203:Reproducibility of Results; D003313:Zea mays / Q000737:chemistry; D015025:Zearalenone / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D004968:Estrogens, Non-Steroidal; D015025:Zearalenone