Development of a real-time immuno-PCR assay for the quantification of 17β-estradiol in water.
Gaudet, D.A., Nilsson, D., Lohr, T., and Sheedy, C. (2015). "Development of a real-time immuno-PCR assay for the quantification of 17β-estradiol in water.", Journal of Environmental Science and Health, Part B: Pesticides, Food Contaminants, and Agricultural Wastes, 50(10), pp. 683-690. doi : 10.1080/03601234.2015.1048097 Access to full text
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−1 to 10 µg mL−1. The RT-iPCR assay's limit of detection was 0.7 pg mL−1. 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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