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Clinical Chemistry 36: 2036-2041, 1990;
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Clinical Chemistry, Vol 36, 2036-2041, Copyright © 1990 by American Association for Clinical Chemistry

Direct chemiluminescence immunoassay of estradiol in saliva

J De Boever, F Kohen, J Bouve, D Leyseele and D VandeKerckhove
Department of Obstetrics and Gynecology, University Hospital, Gent, Belgium.

A sensitive and simple direct solid-phase chemiluminescence immunoassay is described for estradiol in saliva. In this assay, a second antibody is bound to the wells of microtiter plates. Either buffer with standards or saliva (100 microL) is incubated in these wells with monoclonal anti-estradiol antibody and with estradiol-isoluminol conjugate. Incubation time is 2 h. Chemiluminescence of the bound fraction is measured in a manually operated luminometer (Biocounter). The assay has a detection limit of 3.8 pmol/L; analytical recovery of added estradiol is 96.8% (SD 7.0%); within- and between-assay CVs range between 2.5% and 12.7%. Forty unknown saliva samples can be assayed and results calculated within 4.5 h. Results of a slightly modified procedure-with black microtiter plates and a prototype of an automated plate reader (Luminoskan)--compare well with those of the described method (r = 0.97). Because steroid-binding globulins have been found in saliva, the effect of displacing agents on the results of the direct chemiluminescence assay is described.


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K. Tamate, M. Charleton, J. P. Gosling, D. Egan, M. Ishikawa, P. F. Fottrell, and M. M. Kane
Direct colorimetric monoclonal antibody enzyme immunoassay for estradiol-17{beta} in saliva
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