Enzymeimmunoassay of unconjugated estriol in serum and saliva during pregnancy.

An enzymeimmunoassay (EIA) for unconjugated estriol was developed. Estriol-6-CMO-BSA was used as antigen for the antiserum production with low cross reactivities to estrone (0.3%) and estradiol (0.17%). The enzyme conjugate (estriol-peroxidase) was prepared using the mixed anhydride reaction. The antibody bound estriol fraction was separated from the free fraction with the double antibody technique. The test requires a simple photometer for the measurement of enzyme label. The developed estriol-EIA needs a short incubation time (2 hrs). The normal values of estriol in serum were determined. The results are in good agreement with the results of RIA. Furthermore, we found an inverse diurnal rhythm of serum and salivary estriol in comparison to the cortisol level. Low estriol levels in the morning and high estriol concentration in the evening are indices of fetal health. Decreased estriol levels in the morning of several days were found in pathological pregnancies (intrauterine fetal death, fetal distress, hepatose).

A sensitive direct enzyme immunoassay for cortisol in plasma and saliva.

A sensitive enzyme immunoassay for cortisol in plasma and saliva without extraction has been developed. An antibody was raised in rabbits against cortisol-3-CMO-BSA and compared with anti-cortisol-21-HS-HSA. The enzyme label was a covalently linked cortisol-horseradish peroxidase conjugate. Solvent extraction was avoided by use of heat denaturation of transcortin and dilution of plasma 1:300. This assay had a limit of detection of 3 fmol/tube and satisfied the standard criteria of accuracy and precision. The test requires only a simple photometer. The normal values of cortisol in plasma and saliva before and after ACTH, dexamethasone or metyrapone application were determined. Saliva, being easily collected by a noninvasive, stress-free technique is an alternative to plasma. The cortisol concentrations were significantly increased in plasma and saliva of patients with Cushing's syndrome and decreased in patients with Addison's disease.

A simple solid phase enzyme immunoassay for aldosterone in plasma and saliva.

A sensitive, solid phase enzyme immunoassay for the determination of aldosterone in plasma and saliva has been developed. For this purpose anti-aldosterone coated tubes were used. The enzyme marker was a covalently linked aldosterone/horseradish peroxidase conjugate. The assay had a limit of detection of 5 pg/assay tube. The test requires neither centrifuge nor gamma or beta counter, but only a simple photometer. The free/bound separation is based on the solid phase technique with a simple washing step. The normal values for aldosterone in plasma and saliva before and after ACTH application were determined. Saliva, being easily collected by a non-invasive, stress-free technique, is an alternative to plasma. The aldosterone levels in patients with Conn's syndrome were significantly increased in plasma and also in saliva.

Enzyme immunoassay of 17-hydroxyprogesterone in plasma, microfilter paper blood and saliva of newborns, children and patients with congenital adrenal hyperplasia.

An enzyme immunoassay for the measurement of 17-hydroxyprogesterone (OHP) is described. The antigens, OHP-3-CMO-BSA and 11-deoxycortisol-21-hemisuccinate-BSA have been used to produce antisera in rabbits. The enzyme marker was horseradish peroxidase. Bound/free separation was achieved by the double antibody/polyethylene glycol method. The method was used for plasma samples, capillary blood dried on paper disc and for saliva. The normal values for OHP in newborns and children were determined. This method makes possible a selected screening program for congenital adrenal hyperplasia in the newborns.

[Demonstration of a new analytical grading system for the determination of pheochromocytoma within the scope of hypertension control]. / Darstellung eines neuen analytischen Stufensystems zur Erfassung des Phäochromozytoms im Rahmen der Hypertoniebekämpfung.

The problems of the phaeochromocytoma are in its rareness, its appearance--every patient with hypertension can be a carrier of a phaeochromocytoma--, its localisation--10% are situated outside the adrenal glands--, its operability with following permanent normalisation of the pressure. As hypertension is very frequent and the diagnostics is very expensive the finding of carriers of phaeochromocytomas is possible only by means of screening methods. Thus cases with an urgent suspicion are found. Such a method is described, the basis of which is a semiquantitative determination of the vanillic amygdalic acid. Instead of the usually used diazocolour-reagent in connection with a non-optimal buffer a complete nitrophenylantiazotate with a buffer of high ionic strength is taken. The pH-value 10 of the urine which is to be demanded for the colour reaction is thus reached in every case. In order to avoid falsely normal results with every urinary test a preparation with added vanillic amygdalic acid is performed and thus the presence of colour inhibitors is recognized. If the result is pathological a quantitative determination is necessary.

[A specific radioimmunologi determination of plasma estradiol without chromatography during the cycle and in pregnancy and determination of the free non-protein-bound fraction using dialysis]. / Eine spezifische, radioimmunologische Bestimmung des Plasmaöstradiols ohne Chromatographie im Zyklus und in der Schwangerschaft und die Bestimmung des freien, nichtproteingebundenen Anteils mittels Dialyse

The plasma concentrations of estradiol were measured by radioimmunoassay without chromatography. The antibody against estradiol was produced in rabbits immunized with 17beta-estra-diol-6-(0-carboxymethyloxime): bovine-serum-albumin. The lower limit of quantitation is 2.6 pg estradiol. A coefficient of variation of 5.9% was ascertained. The recovery of this method is 93% in the range of 89.8 to 95.2%. The concentrations of estradiol during the menstrual cycle are from 30 pg/ml to 300 pg/ml. Estradiol levels during the anovulatory cycle were found below 70 pg/ml. Similar values (2.0 to 2.5%) of percent of unbound estradiol in normal and anovulatory cycle were calculated. The concentrations of estradiol increase in pregnancy on an average of 16 ng/ml. The percentage of free estradiol decreases (up to the 24 weeks of pregnancy) from 2.3% to 1.2%.