Cornell Research Foundation Inc.

Other

Maria I New

Songja Pang

Bo Dupont

Perrin C White

A cDNA clone is isolated encoding a bovine adrenal cytochrome P-450 specific for steroid 21-hydroxylation (P-450.sub.C21). This plasmid pC21a contains an insert of 520 base pairs. It hybridizes with mRNA encoding P-450.sub.C21. The peptide encoded by the insert is highly homologous to two peptides isolated from porcine P-450.sub.C21 and shows limited homology to the P-450 induced by phenobarbital in rat liver. This probe and its human equivalents can be used to screen in utero i.e. pre-natal as well as post-natal patients for adrenal hyperplasia since it reacts well with human DNA as shown. These probes can also be useful for an industrial fermentation process to produce cortisone in quantity.

Genetic probe used in the detection of adrenal hyperplasia

A method for determination of a steroid such as dehydroepiandrosterone sulfate (DS) in a sample of a human body liquid wherein the liquid sample is transferred to a sheet of microfilter paper and dried before being treated with an aqueous solvent to obtain a mixture wherein the dried body liquid is substantially redissolved in the aqueous solution. The mixture is contacted with an aqueous solution of an agent capable of selectively binding the steroid in the presence of a radioisotopically labeled form of steriod whereby part of the labeled steroid and part of the unlabeled steroid present in the sample are bound by forming a complex with the binding agent. Bound steroids are separated from unbound steroids in the aqueous solution and the radioactivity of at least the separated binding agent-steroids-complex or the unbound steroids is performed to determine the concentration of the hormone as a function of measured radioactivity. Additionally, a means for performing the method is disclosed.

Method for determining steroids in human body liquids

A micromethod for the determination of steroids in human body liquids, in particular blood, is disclosed. A small sample of the liquid is transferred to a sheet of absorbing material, e.g., filter paper. A disc of the impregnated paper, containing the dried sample, in eluted with an aqueous solvent, e.g., a buffer solution. Without separating the paper from the elute, the latter is extracted with an organic solvent. A steroid-containing residue is recovered from the organic extract and is subjected to a conventional radioimmunoassay. The method is particularly suited for the detection of disorders with elevated steroid levels such as congenital adrenal hyperplasia in newborn infants by determining the 17.alpha.-hydroxy-progesterone concentration in small blood samples.

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Maria I New