Urinary estrone conjugate and pregnanediol 3-glucuronide enzyme immunoassays for population research.
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.
O'Connor
Kathleen A
KA
Department of Anthropology and Center for Studies in Demography and Ecology, University of Washington, Seattle WA 98195, USA. oconnork@u.washington.edu
Brindle
Eleanor
E
Holman
Darryl J
DJ
Klein
Nancy A
NA
Soules
Michael R
MR
Campbell
Kenneth L
KL
Kohen
Fortüne
F
Munro
Coralie J
CJ
Shofer
Jane B
JB
Lasley
Bill L
BL
Wood
James W
JW
D000328:Adult; D001459:Bangladesh; D004958:Estradiol / Q000031:analogs & derivatives* / Q000652:urine; D004964:Estriol / Q000031:analogs & derivatives* / Q000652:urine; D004966:Estrogens, Conjugated (USP) / Q000652:urine*; D004970:Estrone / Q000031:analogs & derivatives* / Q000652:urine; D005260:Female; D015200:Fluoroimmunoassay; D006801:Humans; D007124:Immunoenzyme Techniques; D008403:Mass Screening / Q000379:methods*; D008875:Middle Aged; D011276:Pregnanediol / Q000031:analogs & derivatives* / Q000652:urine*; D013048:Specimen Handling; D014481:United States
D004966:Estrogens, Conjugated (USP); C025482:estradiol-3-glucuronide; C022270:pregnanediol-3 alpha-glucuronide; C022810:estriol 3-glucuronide; D004970:Estrone; D004958:Estradiol; D004964:Estriol; D011276:Pregnanediol; C017296:estrone sulfate