Fluorescent PCR and automated fragment analysis in preimplantaion gentic diagnosis for 21-hydroxylase deficiency in congenital adrenal hyperplasia.

Onderzoeksoutput: Articlepeer review

31 Citaten (Scopus)

Samenvatting

Congenital adrenal hyperplasia (CAH) is an autosomal recessive disease which is most often caused by a deficiency in steroid 21-hydroxylase. The disease is characterized by a range of impaired adrenal cortisol and aldosterone synthesis combined with an increased androgen synthesis. These metabolic abnormalities lead to an inability to conserve sodium and virilization of females. The most common mutation causing the severe form of CAH is a conversion of an A or C at nucleotide (nt) 656 to a G in the second intron of the steroid 21-hydroxylase gene (CYP21) causing aberrant splicing of mRNA. A couple was referred to our centre for preimplantation genetic diagnosis (PGD) for 21-hydroxylase deficiency in CAH. A PGD was set up to detect the nt656 A/C-->G mutation using fluorescent polymerase chain reaction (PCR) and subsequent restriction enzyme digestion and fragment analysis on an automated sequencer. Using DNA or single cells from the father, the normal allele could not be amplified. Non-amplification of the normal allele has been previously described in asymptomatic carriers, therefore the PCR was further developed using heterozygous lymphoblasts from the mother. The PCR was shown to be highly efficient (96% amplification), accurate (0% contamination) and reliable (0% allelic drop-out). The couple started PGD treatment and the second PGD cycle resulted in a twin pregnancy. The genotype of the fetuses was determined in our laboratory using chorionic villus sampling material using the method described here. Both fetuses were shown to be heterozygous carriers of the mutation, and two healthy girls were born.
Originele taal-2English
Pagina's (van-tot)691-696
Aantal pagina's6
TijdschriftMolecular Human Reproduction
Volume5
Nummer van het tijdschriftJuly
StatusPublished - jul 1999

Bibliografische nota

Mol Hum reprod.5: 691-696

Vingerafdruk

Duik in de onderzoeksthema's van 'Fluorescent PCR and automated fragment analysis in preimplantaion gentic diagnosis for 21-hydroxylase deficiency in congenital adrenal hyperplasia.'. Samen vormen ze een unieke vingerafdruk.

Citeer dit