Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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One-Step Simultaneous Immunochromatographic Strip Test for Multianalysis of Ochratoxin A and Zearalenone

  • Shim, Won-Bo (Division of Applied Life Science (Brain Korea 21 Program), Graduate School of Gyeongsang National University) ;
  • Dzantiev, Boris B. (A.N.Bach Institute of Biochemistry, Russian Academy of Sciences) ;
  • Eremin, Sergei A. (Department of Chemical Enzymology, Faculty of Chemistry, M. V. Lomonosov Moscow State University) ;
  • Chung, Duck-Hwa (Division of Applied Life Science (Brain Korea 21 Program), Graduate School of Gyeongsang National University)
  • Received : 2008.02.04
  • Accepted : 2008.06.16
  • Published : 2009.01.31
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Abstract

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, respectively, 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 $20/30\;{\mu}g/kg$). The cut-off values of the OS-ICG for the spiked corn were 5 and $10\;{\mu}g/kg$ 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.

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