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Clinical Chemistry 49: 1139-1148, 2003; 10.1373/49.7.1139
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(Clinical Chemistry. 2003;49:1139-1148.)
© 2003 American Association for Clinical Chemistry, Inc.

Urinary Estrone Conjugate and Pregnanediol 3-Glucuronide Enzyme Immunoassays for Population Research

Kathleen A. O’Connor1,a, Eleanor Brindle1, Darryl J. Holman1, Nancy A. Klein2, Michael R. Soules2, Kenneth L. Campbell3, Fortüne Kohen4, Coralie J. Munro5, Jane B. Shofer6, Bill L. Lasley7 and James W. Wood8

1 Department of Anthropology and Center for Studies in Demography and Ecology,
2 Department of Obstetrics and Gynecology, and
6 Department of Medicine, Division of Metabolism, Endocrinology, and Nutrition, University of Washington, Seattle WA 98195.

3 Department of Biology, University of Massachusetts, Boston, MA 02125.

4 Department of Biological Regulation, Weizmann Institute of Science, Rehovet 76100, Israel.
Departments of
5 Population Health and Reproduction and
7 Obstetrics and Gynecology, University of California, Davis, CA 95616.

8 Department of Anthropology and Population Research Institute, Pennsylvania State University, University Park, PA 16802.

aAddress correspondence to this author at: Department of Anthropology, Box 353100, University of Washington, Seattle, WA 98195. Fax 206-543-3285; e-mail oconnork{at}u.washington.edu.

Background: Monitoring of reproductive steroid hormones at the population level requires frequent measurements, hormones or metabolites that remain stable under less than ideal collection and storage conditions, a long-term supply of antibodies, and assays useful for a range of populations. We developed enzyme immunoassays for urinary pregnanediol 3-glucuronide (PDG) and estrone conjugates (E1Cs) that meet these criteria.

Methods: Enzyme immunoassays based on monoclonal antibodies were evaluated for specificity, detection limit, parallelism, recovery, and imprecision. Paired urine and serum specimens were analyzed throughout menstrual cycles of 30 US women. Assay application in different populations was examined with 23 US and 42 Bangladeshi specimens. Metabolite stability in urine was evaluated for 0–8 days at room temperature and for 0–10 freeze-thaw cycles.

Results: Recoveries were 108% for the PDG assay and 105% for the E1C assay. Serially diluted specimens exhibited parallelism with calibration curves in both assays. Inter- and intraassay CVs were <11%. Urinary and serum concentrations were highly correlated: r = 0.93 for E1C–estradiol; r = 0.98 for PDG–progesterone. All Bangladeshi and US specimens were above detection limits (PDG, 21 nmol/L; E1C, 0.27 nmol/L). Bangladeshi women had lower follicular phase PDG and lower luteal phase PDG and E1Cs than US women. Stability experiments showed a maximum decrease in concentration for each metabolite of <4% per day at room temperature and no significant decrease associated with number of freeze-thaw cycles.

Conclusions: These enzyme immunoassays can be used for the field conditions and population variation in hormone metabolite concentrations encountered in cross-cultural research.




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