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

• Published in: Journal of environmental science and health. Part B, Pesticides, food contaminants, and agricultural wastes Pesticides, food contaminants, and agricultural wastes, 2015-10, Vol.50 (10), p.683-690
• Main Authors: Gaudet, Daniel, Nilsson, Denise, Lohr, Tanner, Sheedy, Claudia
• Format: Article
• Language: English
• Subjects: agriculture; antibodies; Biological Assay; Enzyme-Linked Immunosorbent Assay - methods; Estradiol - analysis; Estrogens - analysis; immuno-PCR; Manure; Manure-borne estrogens; Real-Time Polymerase Chain Reaction; Water - chemistry; water quality
• ISSN: 0360-1234; 1532-4109
• DOI: 10.1080/03601234.2015.1048097

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.