Technical in-depth comparison of two massive parallel DNA-sequencing methods for formalin-fixed paraffin-embedded tissue from victims of sudden cardiac death

Whole exome sequencing of FFPE samples—expanding the horizon of forensic molecular autopsies

“…High concordance and lack of artefacts might in part be explained by the comparatively high and uniform coverage obtained in the FFPE samples. Hybridization capture has previously been shown to yield very uniform sequencing compared to amplicon-based protocols [ 23 , 54 ].…”

“…The Twist protocol adopted for FFPE samples using UMIs and Westburg library preparation w/wo enzymatic fragmentation steps has previously been validated for SCD using the cardio diagnostic gene panel CDGP. In short, depending on the degree of fragmentation, the FFPE material was either fragmented further or just end-repaired prior to library preparation where UMI probes were incorporated, followed by pooling and hybridization with Twist ds DNA core exome probes with Twist Human Ref Seq spiked-in [ 23 ].…”

“…In the first tier, a previously published cardio diagnostic gene panel (CDGP) was used. The panel encompasses 84 genes correlated to cardiac diseases and has been used extensively for cardiomyopathies, arrhythmias, and SCD in both forensic and clinical genetics [ 23 , 31 ]. In the second tier, a larger sudden death gene panel (SDGP) was constructed to complement and expand the CGDP.…”

Oxycodone in Forensic Toxicology : Analytical Strategies and Interpretation

“…During the last decade, new powerful techniques have emerged that make whole genome sequencing possible in routine work, by massive parallel sequencing (MPS) (78,90). A recent application in forensic and clinical casework is genotyping in suspected sudden cardiac deaths (90,91). Adolfsson et al describes a methodology based on a cardio diagnostic gene panel of 81 genes connected to several diseases including variants of cardiomyopathy and QT syndromes (91).…”

“…A recent application in forensic and clinical casework is genotyping in suspected sudden cardiac deaths (90,91). Adolfsson et al describes a methodology based on a cardio diagnostic gene panel of 81 genes connected to several diseases including variants of cardiomyopathy and QT syndromes (91). Advantages of full-gene haplotype analysis with MPS in order to predict CYP2D6 metabolizing phenotype was identified by Wendt et al, that concluded an underestimation of poor and intermediate metabolizers using targeted SNPs analysis (92).…”