HiFi long-read RNA sequencing enhances clinical diagnostics in rare disorders

N/A (splicing variants, 25-patient cohort)

PacBio Revio + Kinnex protocol + Iso-Seq/Pigeon workflow (HiFi LR RNA-seq) vs short-read RNA-seq in 25 UK/Netherlands patients with suspected splicing variants: capture of known splicing events and revelation of additional events (intron retention, multiple exon skipping, leaky splicing, variant phasing on RNA, isoform switching)

Comparison of PacBio HiFi long-read RNA-seq (Revio platform, Kinnex + Iso-Seq/Pigeon protocol) to short-read RNA-seq in 25 patients with suspected splicing variants (UK and Netherlands). LR RNA-seq confirms 21 known splicing events and reveals additional effects in 8 cases: intron retention, multiple exon skipping, leaky splicing, variant phasing on the same RNA molecule, and isoform switching. Transcript abundance concordance between approaches is strong (r=0.86 in blood).

Synthesis written by Geno'X. For the full original abstract, please refer to the source publication.

Adding 8 diagnoses in 25 patients is clinically substantial (32% gain). Variant phasing on RNA is a unique long-read advantage inaccessible to short reads. The Kinnex + Iso-Seq pipeline is reproducible and potentially integrable in routine diagnostics for unresolved WGS cases with candidate splicing variants.

EJHG (top human genetics) +2; +8 diagnoses in 25 patients (major additional yield) +3; variant phasing on RNA (unique to LR) +2; reproducible Kinnex pipeline +2