Enzyme-Linked Immunosorbent Assays for Rapid and Quantitative Detection of Insecticidal Crystal Proteins of BT Pesticides

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  • TAKAHASHI Yoshiyuki
    Research Institute of Japan Plant Protection Association
  • HORI Hidetaka
    Biopesticide Laboratory at Advanced Technology Institute, Kubota Corp. Laboratories of Molecular Life Sciences, Graduate School of Science and Technology, Niigata University
  • FURUNO Hidekazu
    Research Institute of Japan Plant Protection Association
  • KAWANO Toshiro
    Research Institute of Japan Plant Protection Association
  • TAKAHASHI Makoto
    Biopesticide Laboratory at Advanced Technology Institute, Kubota Corp.
  • WADA Yutaka
    Research Institute of Japan Plant Protection Association

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Other Title
  • BT剤の殺虫性結晶蛋白のELISA法による迅速・定量検出
  • Enzyme-Linked Immunosorbent Assays for

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Description

Three enzyme-linked immunosorbent assay (ELISA) procedures i. e., one indirect ELISA applied avidin-biotin complex system (ABC-ELISA) and two direct ELISA, antigen captured (AC-ELISA) and double antibody sandwich (DAS-ELISA), were employed to rapidly and quantitatively detect Insecticidal crystal proteins (ICPs) from six commercially available BT pesticides by using antiserum against ICPs of Bacillus thuringiensis subsp. kurstaki HD-1. Assay samples treated with 0.1M NaOH to dissolve ICPs reacted in ELISA more strongly than intact or neutralized ones. The order of sensitivity between the procedures was: ABC-ELISA=DAS-ELISA>AC-ELISA. Reciprocal dilution end points of the BT pesticides detected by ABC-ELISA and DAS-ELISA were 105-107 times and the maximum point (107 times) was roughly equal to 4-8ng/ml of ICPs. ABC-ELISA and AC-ELISA showed non-specific reactions to the samples containing the crude sap of homogenized cabbage, though DAS-ELISA did not show any non-specific reactions. BT pesticide residues on cherry fruits were analyzed by ABC-ELISA and bioassay using larvae of diamondback moth. Based on the results, estimations of the residues were almost the same in both methods.

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