Method for detecting pesticides at the picogram level

G - Physics – 01 – N

Patent

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G01N 33/543 (2006.01) G01N 33/53 (2006.01)

Patent

CA 2067342

2067342 9105259 PCTABS00004 An improved antigen-capture enzyme-linked immunosorbent assay, for measuring the presence of a target pesticide in a medium, comprising the steps of: (a) forming a complex of the pesticide in the medium with an excess of a first antibody of known titer; (b) binding the free antibody from Step (a) to a coating conjugate that is bound to a solid phase; (c) binding a signal-generating labeled antibody to the antibody-coating conjugate complex of Step (b); and (d) determining the amount of pesticide in the medium by comparing the signal generated by the labeled antibody of Step (c) to the signal generated by running the above steps with a sample containing no pesticide and one or more samples containing known concentrations of pesticide; wherein the improvement comprises: (i) diluting the titer of the first antibody employed in Step (a) such that the concentration of pesticide needed to reduce the signal generated by the label in Step (d) by 50 % is less than one-half the concentration required without the dilution; and (ii) increasing the titer of the labeled antibody employed in Step (c) so that the total time necessary to run the assay is about the same as a normal assay without the dilution of (i); thereby (iii) increasing by a factor of at least two the sensitivity of the assay to detect target pesticide.

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