METHOD OF INDIRECT COMPETITIVE IMMUNE-ENZYME ANALYSIS FOR DETERMINING THE CONTENT OF TETRODOTOXIN AND ITS ANALOGUES IN EXTRACTS OF MARINE ORGANISMS

摘要

FIELD: biochemistry.;SUBSTANCE: invention relates to the field of biochemistry, namely, to a method for performing indirect competitive immune-enzyme analysis to determine the content of tetrodotoxin (TTX), 4-epiTTX, 4.9-anhydroTTX, 5-deoxiTTX, 11-deoxiTTX and 5,6,11-trideoxiTTX in extracts of marine organisms. According to this method, a tablet is sensitized by incubation with a solution of bovine serum albumin conjugate with TTX in a carbonate buffer, followed by washing and incubation in a blocking buffer; extracts of marine organisms and a solution of polyclonal antibodies, got by immunization of mice with phytohemagglutinin conjugate with TTX, are added to the wells, followed by incubation and washing of the tablet; a conjugate of secondary antibodies with horseradish peroxidase is added, followed by incubation and washing, orthophenylenediamine is added as a substrate; the solution is kept, and then the reaction is stopped and the optical density is measured at a wavelength of 492 nm.;EFFECT: technical result is to reduce the duration of the analysis of the content of tetrodotoxin with the expansion of the range of detectable analogues, simplify the process of preparing the conjugate for immunization, and eliminate the cross-reaction between the obtained antibodies and the components of the studied extracts.;6 cl, 3 ex, 1 dwg