Prenatal molecular diagnosis of four fetuses at high risk for X-linked adrenoleukodystrophy

Abstract

To investigate methods for prenatal molecular diagnosis of fetuses at high risk for X-linked adrenoleukodystrophy (X-ALD). The amniotic fluid was obtained and genomic DNA was isolated from amniotic fluid cells. Maternal contamination was evaluated by paternity test. PCR-RFLP, sequencing and denaturing high performance liquid chromatography (DHPLC) were used to detect the ABCD1 gene of fetal genome. In the pedigree 1, the PCR product (799 bp) of the fetus 1 and her father (normal control) could be digested with BcnI. No P560L mutation, which was present in the index patient, was detected in the ABCD1 gene from the genomic DNA of the fetus 1 using direct sequencing. In the pedigree 2, the PCR product (232 bp) of the fetus 2 and her father could not be digested with MaeI and no Q177X mutation, which was present in the propositus, was detected in the ABCD1 gene from the genomic DNA of the fetus 2 using direct sequencing. In the pedigree 3, the PCR product (271 bp) was digested with AciI, the pattern of the fetus 3 and the propositus being the same, and the R617C mutation was found in the ABCD1 gene from the genomic DNA of the fetus 3 using direct sequencing. In the pedigree 4, the PCR product (269 bp) was analyzed with the DHPLC, and the pattern of elution peaks of the fetus 4 and her father was similar, but different from that of the propositus. No K276E mutation was detectable in the ABCD1 gene from the genomic DNA of the fetus 4 by using direct sequencing. Judging from the sex of the fetuses, fetuses 1 and 2 were normal homozygotes, fetus 3 was an ALD hemizygote, and fetus 4 was a normal hemizygote. A new protocol for X-ALD prenatal molecular diagnosis is proposed, which would ensure the accuracy of prenatal diagnosis.