Open AccessAccurate genetic diagnosis of Finnish pulmonary arterial hypertension patients using oligonucleotide‐selective sequencing
Author(s) -
Vattulainen Sanna,
Aho Joonas,
Salmenperä Pertteli,
Bruce Siina,
Tallila Jonna,
Gentile Massimiliano,
Sankelo Marja,
Laitinen Tarja,
Koskenvuo Juha W.,
Alastalo TeroPekka,
Myllykangas Samuel
Publication year - 2015
Publication title -
molecular genetics and genomic medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.765
H-Index - 29
ISSN - 2324-9269
DOI - 10.1002/mgg3.147
Subject(s) - sanger sequencing , dna sequencing , gene , genetics , massive parallel sequencing , exome sequencing , oligonucleotide , mutation , biology , medicine , bioinformatics , computational biology
The genetic basis of pulmonary arterial hypertension ( PAH ) among Finnish PAH patients is poorly understood. We adopted a novel‐targeted next‐generation sequencing ( NGS ) approach called Oligonucleotide‐Selective Sequencing ( OS ‐Seq) and developed a custom data analysis and interpretation pipeline to identify pathogenic base substitutions, insertions, and deletions in seven genes associated with PAH ( BMPR 2 , BMPR 1B , ACVRL 1 , ENG , SMAD 9 , CAV 1 , and KCNK 3 ) from Finnish PAH patients. This study represents the first clinical study with OS ‐Seq technology on patients suffering from a rare genetic disorder. We analyzed DNA samples from 21 Finnish PAH patients, whose BMPR 2 and ACVRL 1 mutation status had been previously studied using Sanger sequencing. Our sequencing panel covered 100% of the targeted base pairs with >15× sequencing depth. Pathogenic base substitutions were identified in the BMPR 2 gene in 29% of the Finnish PAH cases. Two of the pathogenic variant‐positive patients had been previously tested negative using Sanger sequencing. No clinically significant variants were identified in the six other PAH genes. Our study validates the use of targeted OS ‐Seq for genetic diagnostics of PAH and revealed pathogenic variants that had been previously missed using Sanger sequencing.