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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or quantified.

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ELIME (Enzyme Linked Immuno Magnetic Electrochemical) Method for Mycotoxin Detection
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ELIME (Enzyme Linked Immuno Magnetic Electrochemical) Method for Mycotoxin Detection

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Enzyme-linked immunosorbent assay for the pyrethroid deltamethrin.

Hu-Jang Lee1, Guomin Shan, Takaho Watanabe

  • 1Department of Entomology and Cancer Research Center, University of California, Davis 95616, USA.

Journal of Agricultural and Food Chemistry
|September 19, 2002
PubMed
Summary

A new competitive enzyme-linked immunosorbent assay (ELISA) was developed for detecting deltamethrin. This sensitive method offers low detection limits and minimal cross-reactivity with other pyrethroids, making it suitable for environmental monitoring.

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Area of Science:

  • Analytical Chemistry
  • Environmental Science
  • Immunochemistry

Background:

  • Deltamethrin is a widely used synthetic pyrethroid insecticide.
  • Accurate and sensitive detection methods are crucial for environmental monitoring and risk assessment.
  • Existing detection methods may lack specificity or require complex sample preparation.

Purpose of the Study:

  • To develop and optimize a sensitive competitive enzyme-linked immunosorbent assay (ELISA) for deltamethrin detection.
  • To characterize the assay's performance, including sensitivity, specificity, and optimal conditions.
  • To evaluate the applicability of the developed ELISA for analyzing deltamethrin in river water samples.

Main Methods:

  • Synthesis and conjugation of two haptens with thyroglobulin to generate immunogens.
  • Generation and screening of antisera against various coating antigens to identify the most effective antibody.
  • Optimization of assay conditions, including organic cosolvent, pH, and ionic strength.
  • Integration of C(18) sorbent-based solid-phase extraction for sample pre-concentration.
  • Validation of the method using fortified river water samples.

Main Results:

  • The developed competitive ELISA demonstrated high sensitivity for deltamethrin detection, with an I(50) of 17.5 ± 3.6 µg/L and a lower detection limit of 1.1 ± 0.5 µg/L.
  • The assay exhibited low cross-reactivity with other common pyrethroids, indicating good specificity.
  • Methanol at 40% (v/v) was identified as the optimal organic cosolvent, and the assay was stable within a pH range of 5.0–8.0.
  • Solid-phase extraction enhanced the overall method's sensitivity for river water samples, yielding good recoveries and correlation with spike levels.

Conclusions:

  • A sensitive and specific competitive ELISA for deltamethrin detection has been successfully developed.
  • The assay is robust, with optimal performance under specific solvent, pH, and ionic strength conditions.
  • The integrated method, including solid-phase extraction, is effective for the analysis of deltamethrin in environmental water samples.