Open AccessIdentification and validation of loss of function variants in clinical contexts
Author(s) -
Lescai Francesco,
Marasco Elena,
Bacchelli Chiara,
Stanier Philip,
Mantovani Vilma,
Beales Philip
Publication year - 2014
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/mgg3.42
Subject(s) - indel , pipeline (software) , context (archaeology) , computer science , identification (biology) , computational biology , snp , queue , single nucleotide polymorphism , exome sequencing , loss function , genetics , biology , gene , mutation , phenotype , paleontology , botany , genotype , programming language
The choice of an appropriate variant calling pipeline for exome sequencing data is becoming increasingly more important in translational medicine projects and clinical contexts. Within GOS gene, which facilitates genetic analysis as part of a joint effort of the University College London and the Great Ormond Street Hospital, we aimed to optimize a variant calling pipeline suitable for our clinical context. We implemented the GATK /Queue framework and evaluated the performance of its two callers: the classical UnifiedGenotyper and the new variant discovery tool HaplotypeCaller. We performed an experimental validation of the loss‐of‐function (LoF) variants called by the two methods using Sequenom technology. UnifiedGenotyper showed a total validation rate of 97.6% for LoF single‐nucleotide polymorphisms ( SNP s) and 92.0% for insertions or deletions ( INDEL s), whereas HaplotypeCaller was 91.7% for SNP s and 55.9% for INDEL s. We confirm that GATK /Queue is a reliable pipeline in translational medicine and clinical context. We conclude that in our working environment, UnifiedGenotyper is the caller of choice, being an accurate method, with a high validation rate of error‐prone calls like LoF variants. We finally highlight the importance of experimental validation, especially for INDEL s, as part of a standard pipeline in clinical environments.