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Development of a real-time immuno-PCR assay for the quantification of 17β-estradiol in water

Gaudet, D., Nilsson, D., Lohr, T., Sheedy, C. (2015). Development of a real-time immuno-PCR assay for the quantification of 17β-estradiol in water, 50(10), 683-690. http://dx.doi.org/10.1080/03601234.2015.1048097

Abstract

Copyright © 2015 Crown copyright. A competitive real-time (RT) immuno-polymerase chain reaction (iPCR) (RT-iPCR) assay was developed for the sensitive quantification of 17β-estradiol in water. Using a universal iPCR method and polyclonal antibodies, 17β-estradiol was accurately quantified at concentrations ranging from 1 pg mL<sup>−1</sup> to 10 µg mL<sup>−1</sup>. The RT-iPCR assay's limit of detection was 0.7 pg mL<sup>−1</sup>. The RT-iPCR assay provided an 800-fold increase in sensitivity as well as an expanded working range compared with the corresponding enzyme-linked immunosorbent assay. Assay cross-reactivity to estrone and estriol, two structurally related estrogens, was below 8%. Water samples spiked with 17β-estradiol were analyzed by RT-iPCR to determine the assay's potential as a rapid screen for the monitoring of manure-borne estrogens in the environment. The assay showed recoveries of 82, 102 and 103% for Milli-Q, tap, and irrigation water, respectively, without requiring sample extraction or concentration prior to analysis.

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