Detection of steroid 21-hydroxylase alleles using gene-specific PCR and a multiplexed ligation detection reaction

Journal Article · Fri Sep 01 04:00:00 EDT 1995 · Genomics

DOI:https://doi.org/10.1006/geno.1995.1226· OSTI ID:433296

Day, D J; Barany, F; Speiser, P W ^[1]

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1. Cornell Univ. Medical College, New York, NY (United States); and others

Steroid 21-hydroxylase deficiency is the most common cause of congenital adrenal hyperplasia, an inherited inability to synthesize cortisol that occurs in 1 in 10,000-15,000 births. Affected females are born with ambiguous genitalia, a condition that can be ameliorated by administering dexamethasone to the mother for most of gestation. Prenatal diagnosis is required for accurate treatment of affected females as well as for genetic counseling purposes. Approximately 95% of mutations causing this disorder result from recombinations between the gene encoding the 21-hydroxylase enzyme (CYP21) and a linked, highly homologous pseudogene (CYP21P). Approximately 20% of these mutations are gene deletions, and the remainder are gene conversions that transfer any of nine deleterious mutations from the CYP21P pseudogene to CYP21. We describe a methodology for genetic diagnosis of 21-hydroxylase deficiency that utilizes gene-specific PCR amplification in conjunction with thermostable DNA ligase to discriminate single nucleotide variations in a multiplexed ligation detection assay. The assay has been designed to be used with either fluorescent or radioactive detection of ligation products by electrophoresis on denaturing acrylamide gels and is readily adaptable for use in other disease systems. 30 refs., 5 figs.

• OSTI ID: 433296
• Journal Information: Genomics, Journal Name: Genomics Journal Issue: 1 Vol. 29; ISSN 0888-7543; ISSN GNMCEP
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
BIOASSAY
BIOSYNTHESIS
CONGENITAL DISEASES
DETECTION
DIAGNOSIS
DNA HYBRIDIZATION
EMBRYOS
EVALUATION
GENE MUTATIONS
GENE RECOMBINATION
GENES
GENETIC MAPPING
HISTOCOMPATIBILITY COMPLEX
HUMAN CHROMOSOME 6
HYDROCORTISONE
HYDROXYLASES
METABOLIC DISEASES
POLYMERASE CHAIN REACTION
RADIOENZYMATIC ASSAY
SENSITIVITY
STEROIDS