Fast and Easy Nanopore Sequencing Workflow for Rapid Genetic Testing of Familial Hypercholesterolemia
Familial hypercholesterolemia (FH) is an autosomal dominant lipid metabolism disorder characterized by severely elevated plasma low-density lipoprotein cholesterol levels. The disease is caused by mutations in 3 genes (LDLR, APOB and PCSK9) while over 90% of the mutations are located within the L...
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|Familial hypercholesterolemia (FH) is an autosomal dominant lipid metabolism disorder
characterized by severely elevated plasma low-density lipoprotein cholesterol levels. The
disease is caused by mutations in 3 genes (LDLR, APOB and PCSK9) while over 90% of
the mutations are located within the LDLR gene. Thus, genetic analysis of the LDLR gene is
the first step in the genetic diagnosis of FH. However, conventional methods like Sanger
and NextGen sequencing are still costly and time-consuming. In contrast, Oxford
Nanopore technology sequencing is an emerging third-generation sequencing
technology featured by easy operability, low cost, small size and the capability of
parallel sample sequencing. Here, we present an easy Nanopore-sequencing-based
workflow for the rapid genetic testing of FH taking only 3 days and costing less than
$50 per sample without the requirement for deep bioinformatic knowledge. Using our
workflow, we were able to identify the underlying pathogenic variants of 10 FH patients
including one novel, not yet recorded pathogenic variants. Our workflow allows the rapid
evaluation of the pathogenic variants by utilizing detailed variant information from Ensembl.
Additionally, our workflow is not restricted to sequencing the LDLR gene alone but can be
easily adapted to the other FH-causing genes and more importantly, to any desired gene
contributing to any hereditary disease. Therefore, our workflow is an attractive opportunity
for every diagnostic laboratory to offer fast and easy in-house genetic diagnostics.
|Gefördert durch den Open-Access-Publikationsfonds der UB Marburg.