Mutation analysis in Bardet-Biedl syndrome by DNA pooling and massively parallel resequencing in 105 individuals

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

Research output: Contribution to journalArticlepeer-review


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.
Original languageEnglish
Pages (from-to)79-90
Number of pages12
JournalHuman Genetics
Issue number1
Publication statusPublished - Jan 2011


  • Amino Acid Sequence
  • Bardet-Biedl Syndrome
  • Base Sequence
  • Chromosome Mapping
  • Cohort Studies
  • Consanguinity
  • DNA Mutational Analysis
  • Exons
  • Genetic Heterogeneity
  • Genetic Loci
  • Genetic Predisposition to Disease
  • Group II Chaperonins
  • Humans
  • Microtubule-Associated Proteins
  • Molecular Sequence Data
  • Mutation
  • Proteins


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