Abstract |
Bardet-Biedl syndrome (BBS) is a rare, primarily autosomal-recessive ciliopathy. The phenotype of this pleiotropic disease includes retinitis pigmentosa, postaxial polydactyly, truncal obesity, learning disabilities, hypogonadism and renal anomalies, among others. To date, mutations in 15 genes (BBS1-BBS14, SDCCAG8) have been described to cause BBS. The broad genetic locus heterogeneity renders mutation screening time-consuming and expensive. We applied a strategy of DNA pooling and subsequent massively parallel resequencing (MPR) to screen individuals affected with BBS from 105 families for mutations in 12 known BBS genes. DNA was pooled in 5 pools of 21 individuals each. All 132 coding exons of BBS1-BBS12 were amplified by conventional PCR. Subsequent MPR was performed on an Illumina Genome Analyzer II™ platform. Following mutation identification, the mutation carrier was assigned by CEL I endonuclease heteroduplex screening and confirmed by Sanger sequencing. In 29 out of 105 individuals (28%), both mutated alleles were identified in 10 different BBS genes. A total of 35 different disease-causing mutations were confirmed, of which 18 mutations were novel. In 12 additional families, a total of 12 different single heterozygous changes of uncertain pathogenicity were found. Thus, DNA pooling combined with MPR offers a valuable strategy for mutation analysis of large patient cohorts, especially in genetically heterogeneous diseases such as BBS.
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Authors | Sabine Janssen, Gokul Ramaswami, Erica E Davis, Toby Hurd, Rannar Airik, Jennifer M Kasanuki, Lauren Van Der Kraak, Susan J Allen, Philip L Beales, Nicholas Katsanis, Edgar A Otto, Friedhelm Hildebrandt |
Journal | Human genetics
(Hum Genet)
Vol. 129
Issue 1
Pg. 79-90
(Jan 2011)
ISSN: 1432-1203 [Electronic] Germany |
PMID | 21052717
(Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
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Chemical References |
- Bbs1 protein, human
- Bbs2 protein, human
- Microtubule-Associated Proteins
- Proteins
- Group II Chaperonins
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Topics |
- Amino Acid Sequence
- Bardet-Biedl Syndrome
(genetics)
- Base Sequence
- Chromosome Mapping
- Cohort Studies
- Consanguinity
- DNA Mutational Analysis
(methods)
- Exons
(genetics)
- Genetic Heterogeneity
- Genetic Loci
(genetics)
- Genetic Predisposition to Disease
- Group II Chaperonins
(genetics)
- Humans
- Microtubule-Associated Proteins
- Molecular Sequence Data
- Mutation
(genetics)
- Proteins
(genetics)
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