A few enzymeimmunoassay (EIA) for steroid hormincs cspecially testosterone (T) have been reported but was nt suitable for al biological samples.
The present study was designed to develop a rapid, sensitive EIA and to apply this technique for study th...
A few enzymeimmunoassay (EIA) for steroid hormincs cspecially testosterone (T) have been reported but was nt suitable for al biological samples.
The present study was designed to develop a rapid, sensitive EIA and to apply this technique for study the physiological changes of T in biological samples. Saliva samples were collected on 06:00-19:00 hour during one menstrual cycle from 18 normally menstruating women a and on 10:00-12:00 hour from 20 normal men.
The present study shows an established T-EIA, using horse radish peroxidase (HRP), which was covalently bonded to testosterone-3-carboxy methyl (T-3-CMO). One batch of T-antisera was also covalently linked to microcrystalline cellulose particles by an mixed anhydride method in order to facilitate separation of bound and free steroid. The esta-blished EIA was validated in terms of sensitivity, accuracy, specificity, precisions etc., comparing with conventional radioimmunoassay, which has also been established in the present study. The sensitivity of the established EIA was less than 25pg/tube.
The correlation coefficients between the expexted Tvalues and observed T-values meas-ured by FIA or RIA were r-0.985 and 1-0.941 respectively. The cross reactivity of an tiserum in EIA was a little higher than that of RIA, especilly by 5 alpha-DHT.
The intra-and inter-assay precision of the present FIA were extremely similar to those
of RIA. The present study also demonstrates that the normal T-values in salive of Ko-rean male and femal samples are 265.65+15.80pmol/l and 109.7412. Olpmol/l respe ctively,
The present EIA seems to be established and suitable for use in the endocrinological
studies. The advantages of this EIA system also might make the present T-EIA an ideal procedure for use in a routine steroid laboratory especially in underdeveloping poor country.