This assay was originally developed for the World Health Organization as a non-invasive monitor of ovarian function. It has been widely used in clinical trials and in the assessment of natural family planning methods.

Urine samples are diluted in buffer and the Estrone-3-Glucuronide concentration measured by EIA (enzyme labelled immunoassay with magnetic second antibody separation). All samples, standards and quality control specimens are assayed in duplicate.

Assays are controlled through the use of at least 2 Internal Quality Control samples in every run. In addition the performance of every run is closely monitored for various parameters including imprecision, antibody binding of tracer, non-specific binding and reproducibility of the standard curve.

• Standard Curve: The range of the standard curve is 0 to 10.7 nmol/L.
• Precision: The between-run precision of the assay is generally less than 10% CV.
• Sensitivity: The sensitivity of the assay is approximately 0.1 nmol/L (defined as 2 standard deviations in the measurement of zero dose).

The cross-reactions of the Estrone-3-Glucuronide antibody (estimated as the relative potency causing a 50% displacement of zero binding) are shown below:

Substance % Cross Reaction Estrone-3-Glucuronide 100 Estrone 97 Estrone-3-Sulphate 231

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