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A sensitive enzymeimmunoassay with a fluorimetric end-point for the determination of testosterone in female plasma and saliva


A sensitive enzymeimmunoassay with a fluorimetric end-point for the determination of testosterone in female plasma and saliva



Steroids 35(1): 89-101



ISSN/ISBN: 0039-128X

PMID: 6990557

A fluorimetric enzymeimmunoassay was developed having the sensitivity (500 f[femto]g/assay tube) required for determining testosterone [T] concentrations in female plasma and saliva samples. The assay featured a solid-phase antiserum raised against an 11.alpha.-hydroxytestosterone-11-hemisuccinate bovine serum albumin conjugate, an 11.alpha.-hydroxytestosterone-11-hemisuccinate horseradish peroxidase conjugate as the enzyme label, and p-hydroxyphenylacetic acid as the substrate for the development of fluorescence. Specificity was ensured by extracting T from samples with a solid-phase anti-testosterone-3- O-carboxymethyl -oxime serum. The assay satisfied accepted validation criteria providing results in good agreement with routine radioimmunoassay procedures in both plasma (r [correlation coefficient] > 0.98, n = 28) and saliva (r > 0.99, n = 28). In saliva samples collected at 2 hourly intervals by normal healthy women (n = 5) T concentrations showed a well-defined circadian rhythm; the mean T concentration in early morning samples (174 pmol/l) fell by 83% in late evening collections. In healthy female volunteers (n = 7), mean salivary T concentrations in samples collected daily throughout 1 complete cycle ranged from 50-218 pmol/l. Following dexamethasone administration T concentrations in plasma fell by approximately 50% and salivary concentrations were undetectable after 1 h. This enzymeimmunoassay may be useful in studies of female infertility.

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Accession: 004623477

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