One-step simultaneous immunochromatographic strip test for multianalysis of ochratoxin a and zearalenone.
Individual immunochromatographic assays (ICG) for ochratoxin A (OTA) and zearalenone (ZEA) were optimized and used in the development of a one-step simultaneous immunochromatographic assay (OS-ICG) for the rapid multianalysis of two mycotoxins in corn samples. The nitrocellulose membrane of the OS-ICG was treated with OTA-bovine serum albumin (BSA), ZEA-ovalbumin (OVA), and anti-mouse IgG in the OTA test, ZEA test, and control zones, respectively. Monoclonal antibody-gold conjugates (OTA3 MAb-gold and ZEA2C5 MAb-gold) were sprayed onto the conjugate pad. The visual detection limits were 2.5 and 5 ng/ml for OTA and ZEA, resepectively, and the results were obtained within 15 min after starting the analysis. An efficient, simple, and rapid extraction method using 30% MeOH/PBS was established and validated by analyzing the corn samples spiked with OTA/ZEA mixtures (0/0, 5/10, 10/20, and 20/30 microg/kg). The cut-off values of the OS-ICG for the spiked corn were 5 and 10 microg/kg for OTA and ZEA, respectively. Natural corn samples were analyzed by OS-ICG, direct competitive enzyme-linked immunosorbent assay (DC-ELISA), and HPLC. Results of the OS-ICG were in good agreement with those obtained by DC-ELISA and HPLC. The developed OS-ICG offers a rapid, easy-to-use, and portable analytical system and can be used as a convenient qualitative tool for the on-site simultaneous determination of OTA and ZEA in cereals, food, and agricultural products in one analytical cycle.
Shim
Won-Bo
WB
Division of Applied Life Science (Brain Korea 21 Program), Graduate School of Gyeongsang National University, Chinju, Korea.
Dzantiev
Boris B
BB
Eremin
Sergei A
SA
Chung
Duck-Hwa
DH
D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D002273:Carcinogens / Q000032:analysis; D002845:Chromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D004968:Estrogens, Non-Steroidal / Q000032:analysis; D005504:Food Analysis / Q000379:methods; D017979:Gold Colloid; D007118:Immunoassay / Q000379:methods*; D009793:Ochratoxins / Q000032:analysis* / Q000276:immunology; D011786:Quality Control; D011934:Reagent Strips; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D003313:Zea mays / Q000737:chemistry; D015025:Zearalenone / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D002273:Carcinogens; D004968:Estrogens, Non-Steroidal; D017979:Gold Colloid; D009793:Ochratoxins; D011934:Reagent Strips; C025589:ochratoxin A; D015025:Zearalenone