title: Efficient molecular diagnostic strategy for ABCC6 in pseudoxanthoma elasticum.
abstract: Pseudoxanthoma elasticum (PXE) is a hereditary disorder of connective tissue with skin, cardiovascular, and visual involvement. In familial cases, PXE usually segregates in an autosomal recessive fashion. The aim of this manuscript is to describe an efficient strategy for DNA diagnosis of PXE. The two most frequent mutations, R1141X and an ABCC6 del exons 23-29, as well as a core set of mutations, were identified by restriction enzyme digestion and size separation on agarose gels. Next, in the remaining patient group in which only one or no mutant allele was found, the complete coding sequence was analyzed using denaturing high-performance liquid chromatography (dHPLC). All variations found were confirmed by direct DNA sequencing. Finally, Southern blot was used to investigate the potential presence of small or large deletions. Twenty different mutations, including two novel mutations in the ABCC6 gene, were identified in 80.3% of the 76 patients, and 58.6% of the 152 ABCC6 alleles analyzed. With this strategy, 70 (78.7%) out of 89 mutant alleles could be detected within a week. We conclude that this strategy leads to both reliable and time-saving screening for mutations in the ABCC6 gene in sporadic cases and in families with PXE.
authors.affiliation: Netherlands Ophthalmic Research Institute, KNAW, Amsterdam, The Netherlands.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Hu
authors.forename: Astrid
authors.initials: A
authors.lastname: Plomp
authors.forename: Theo
authors.initials: T
authors.lastname: Gorgels
authors.forename: Jacoline Ten
authors.initials: JT
authors.lastname: Brink
authors.forename: Willem
authors.initials: W
authors.lastname: Loves
authors.forename: Marcel
authors.initials: M
authors.lastname: Mannens
authors.forename: Paulus T V M
authors.initials: PT
authors.lastname: de Jong
authors.forename: Arthur A B
authors.initials: AA
authors.lastname: Bergen
mesh_terms: D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000737:chemistry / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D018703:Founder Effect; D005817:Genetic Counseling; D006801:Humans; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000235:genetics*; D009154:Mutation / Q000235:genetics*; D009691:Nucleic Acid Denaturation; D011561:Pseudoxanthoma Elasticum / Q000175:diagnosis* / Q000235:genetics
chemical_list: D004247:DNA; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C102029:ABCC6 protein, human