Hapten Synthesis for the Development of a Competitive Inhibition Enzyme-Immunoassay for Thiram

Françoise Gueguen (1) , Frédéric Boisdé (2) , Anne-Laurence Queffelec (2, 1) , Jean-Pierre Haelters (2) , Daniel Thouvenot (1) , Bernard Corbel (2) , Patrice Nodet (1)

1 LUBEM - Laboratoire Universitaire de Biodiversité et Ecologie Microbienne
2 CEMCA - Chimie, Electrochimie Moléculaires et Chimie Analytique

An enzyme-linked immunosorbent assay (ELISA) was developed for the fungicide thiram. Two types of haptens were synthesized. The first type exhibits the two symmetrical N-alkyl dithiocarbamate patterns of thiram with a spacer arm linked to one of the N-methyl terminal group. The second type exhibits one of the two symmetrical N-alkyl dithiocarbamate patterns of thiram with a variable-length spacer arm linked to one sulfur atom. Polyclonal antibodies suitable for thiram detection were obtained from immunization with an hapten of the first type, while haptens of the second type were used as coating antigens to develop a competitive ELISA against thiram. The IC50 value for thiram was estimated to be 0.24 mug/mL, with a detection limit of 0.03 mug/mL. The assay seems to be thiram-specific since no or little cross-reaction with other dithiocarbamates were observed.

Mots clés: Thiram; dithiocarbamates; hapten; ELISA

Domaines: Environnement et Société; Alimentation et Nutrition; Biotechnologies; Agronomie; Génie chimique

Françoise Gueguen, Frédéric Boisdé, Anne-Laurence Queffelec, Jean-Pierre Haelters, Daniel Thouvenot, et al.. Hapten Synthesis for the Development of a Competitive Inhibition Enzyme-Immunoassay for Thiram. Journal of Agricultural and Food Chemistry, 2000, 48 (10), pp.4492-4499. ⟨10.1021/jf000378g⟩. ⟨hal-01329487⟩