Urinary estrone conjugate and pregnanediol 3-glucuronide enzyme immunoassays for population research

Kathleen A. O'Connor, Eleanor Brindle, Darryl J. Holman, Nancy A. Klein, Michael R. Soules, Kenneth L. Campbell, Fortüne Kohen, Coralie J. Munro, Jane B. Shofer, Bill L. Lasley, James William Wood

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish (US)
Pages (from-to)1139-1148
Number of pages10
JournalClinical Chemistry
Volume49
Issue number7
DOIs
StatePublished - Jul 1 2003

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Estrone
Immunoenzyme Techniques
Assays
Metabolites
Enzymes
Research
Population
Hormones
Limit of Detection
Steroid hormones
Urine
Recovery
Temperature
Follicular Phase
Luteal Phase
Menstrual Cycle
Serum
Calibration
Progesterone
pregnanediol-3 alpha-glucuronide

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

O'Connor, K. A., Brindle, E., Holman, D. J., Klein, N. A., Soules, M. R., Campbell, K. L., ... Wood, J. W. (2003). Urinary estrone conjugate and pregnanediol 3-glucuronide enzyme immunoassays for population research. Clinical Chemistry, 49(7), 1139-1148. https://doi.org/10.1373/49.7.1139
O'Connor, Kathleen A. ; Brindle, Eleanor ; Holman, Darryl J. ; Klein, Nancy A. ; Soules, Michael R. ; Campbell, Kenneth L. ; Kohen, Fortüne ; Munro, Coralie J. ; Shofer, Jane B. ; Lasley, Bill L. ; Wood, James William. / Urinary estrone conjugate and pregnanediol 3-glucuronide enzyme immunoassays for population research. In: Clinical Chemistry. 2003 ; Vol. 49, No. 7. pp. 1139-1148.
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O'Connor, KA, Brindle, E, Holman, DJ, Klein, NA, Soules, MR, Campbell, KL, Kohen, F, Munro, CJ, Shofer, JB, Lasley, BL & Wood, JW 2003, 'Urinary estrone conjugate and pregnanediol 3-glucuronide enzyme immunoassays for population research', Clinical Chemistry, vol. 49, no. 7, pp. 1139-1148. https://doi.org/10.1373/49.7.1139

Urinary estrone conjugate and pregnanediol 3-glucuronide enzyme immunoassays for population research. / O'Connor, Kathleen A.; Brindle, Eleanor; Holman, Darryl J.; Klein, Nancy A.; Soules, Michael R.; Campbell, Kenneth L.; Kohen, Fortüne; Munro, Coralie J.; Shofer, Jane B.; Lasley, Bill L.; Wood, James William.

In: Clinical Chemistry, Vol. 49, No. 7, 01.07.2003, p. 1139-1148.

Research output: Contribution to journalArticle

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T1 - Urinary estrone conjugate and pregnanediol 3-glucuronide enzyme immunoassays for population research

AU - O'Connor, Kathleen A.

AU - Brindle, Eleanor

AU - Holman, Darryl J.

AU - Klein, Nancy A.

AU - Soules, Michael R.

AU - Campbell, Kenneth L.

AU - Kohen, Fortüne

AU - Munro, Coralie J.

AU - Shofer, Jane B.

AU - Lasley, Bill L.

AU - Wood, James William

PY - 2003/7/1

Y1 - 2003/7/1

N2 - 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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O'Connor KA, Brindle E, Holman DJ, Klein NA, Soules MR, Campbell KL et al. Urinary estrone conjugate and pregnanediol 3-glucuronide enzyme immunoassays for population research. Clinical Chemistry. 2003 Jul 1;49(7):1139-1148. https://doi.org/10.1373/49.7.1139