Linked Life Data

19814614

Source:http://linkedlifedata.com/resource/pubmed/id/19814614

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
2009-10-9
pubmed:abstractText
Dystrophic epidermolysis bullosa is a heritable skin disease in which blisters occur because of a defect in type VII collagen resulting from mutations in the COL7A1 gene that is composed of 118 exons. Although a few mutations are specific to certain populations owing to founder effects, and although a few mutational hotspots exist, most mutations are unique to families and can be found scattered throughout the entire COL7A1 gene. This emphasizes the need for a sensitive, reliable, and efficient mutation scanning technique. Therefore, we developed a conformation-sensitive capillary electrophoresis (CSCE) system for COL7A1 mutation scanning. Here we report on the design and validation of this system. The CSCE technique is based on the principle of heteroduplex formation when polymerase chain reaction-amplified DNA fragments containing heterozygous sequence changes are slowly reannealed. These fluorescently labeled fragments have different migration characteristics and can be detected on a multi-capillary automated sequencer. Validation was performed by analysis of 29 known COL7A1 sequence changes, covering 33% of amplicons. After optimization of the conditions, all 29 sequence changes were detected by the CSCE system, irrespective of length or CG-content of amplicons and position of sequence changes, reflecting an analytical sensitivity of 90.2-100% (95% confidence interval). We conclude that this CSCE system is a rapid, reliable, cost-effective, and highly sensitive way of mutation scanning for COL7A1 in a molecular genetics service laboratory.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
1945-0257
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
13
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
589-97
pubmed:meshHeading
pubmed:year
2009
pubmed:articleTitle
Design and validation of a conformation-sensitive capillary electrophoresis system for mutation identification of the COL7A1 gene with automated peak comparison.
pubmed:affiliation
Department of Genetics, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands. p.c.van.den.akker@medgen.umcg.nl
pubmed:publicationType
Journal Article, Validation Studies