Preimplantation genetic diagnosis of X-linked adrenoleukodystrophy with gender determination using multiple displacement amplification

• Published in: Fertility and sterility Vol. 88; no. 5; pp. 1327 - 1333
• Main Authors: Lledó, Belén, Ph.D, Bernabeu, Rafael, M.D, Ten, Jorge, Ph.D, Galán, Francisco M., Ph.D, Cioffi, Luigi, Ph.D
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01.11.2007; Elsevier Science
• Subjects: Adrenals. Adrenal axis. Renin-angiotensin system (diseases); Adrenoleukodystrophy - diagnosis; Adrenoleukodystrophy - genetics; Adult; ALD; Biological and medical sciences; Endocrinopathies; Errors of metabolism; Female; Genetic Linkage - genetics; Gynecology. Andrology. Obstetrics; Humans; Internal Medicine; Lipids (lysosomal enzyme disorders, storage diseases); Male; MDA; Medical sciences; Metabolic diseases; multiple displacement amplification; Non tumoral diseases. Target tissue resistance. Benign neoplasms; Nucleic Acid Amplification Techniques - methods; Obstetrics and Gynecology; Pedigree; PGD; Polymerase Chain Reaction - methods; Preimplantation Diagnosis - methods; preimplantation genetic diagnosis; Sex Determination Processes; X-linked adrenoleukodystrophy; ALD; MDA; multiple displacement amplification; preimplantation genetic diagnosis; PGD; X-linked adrenoleukodystrophy; Endocrinopathy; Sex linked character; Sex; Lipids; Sphingolipidosis; Enzymopathy; Adrenal gland diseases; Genetics; Degenerative disease; X-Chromosome; Lipoidosis; Nervous system diseases; Multiple; Leukodystrophy; Gynecology; Metabolic diseases; Obstetrics; Displacement; Genetic disease; Cerebral disorder; Amplification; Preimplantation diagnosis; Adrenal insufficiency; Adrenoleukodystrophy; Central nervous system disease
• ISSN: 0015-0282; 1556-5653
• DOI: 10.1016/j.fertnstert.2007.01.034

Objective To evaluate the use of multiple displacement amplification (MDA) for whole genome amplification in the preimplantation genetic diagnosis (PGD) of X-linked adrenoleukodystrophy. Design MDA was used to amplify the whole genome directly from a single blastomere. MDA products were used for polymerase chain reaction (PCR) analysis of two polymorphic markers flanking the ABCD1 gene and a new X/Y marker, X22, to sex embryos in an X-linked adrenoleukodystrophy PGD program. Setting Fertility and gynecology private center in Alicante, Spain. Patient(s) A couple in which the wife is a carrier of the ABCD1 gene mutation (676A→C) that was previously identified in her family. Intervention(s) MDA of single blastomere and PCR tests for PGD. Main Outcome Measure(s) The ability to analyze single blastomeres for X-linked adrenoleukodystrophy using MDA. Result(s) The development of an MDA-PGD protocol for X-linked adrenoleukodystrophy allowed for the diagnosis of five embryos. These were biopsied on day 3 of culture and analyzed. One embryo was an affected male and one embryo was a female carrier. Three healthy female embryos were transferred 48 hours after biopsy. Unfortunately, no pregnancy was achieved. Conclusion(s) The MDA technique is useful for overcoming the problem of insufficient genomic DNA in PGD and allows the simultaneous amplification of different targets to perform a diagnosis of any known gene defect and a sexing test by standard methods and conditions.