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- Title
Performance evaluation of Sanger sequencing for the diagnosis of primary hyperoxaluria and comparison with targeted next generation sequencing.
- Authors
Williams, Emma L.; Bagg, Eleanor A. L.; Mueller, Michael; Vandrovcova, Jana; Aitman, Timothy J.; Rumsby, Gill
- Abstract
Definitive diagnosis of primary hyperoxaluria ( PH) currently utilizes sequential Sanger sequencing of the AGXT, GRPHR, and HOGA1 genes but efficacy is unproven. This analysis is time-consuming, relatively expensive, and delays in diagnosis and inappropriate treatment can occur if not pursued early in the diagnostic work-up. We reviewed testing outcomes of Sanger sequencing in 200 consecutive patient samples referred for analysis. In addition, the Illumina Truseq custom amplicon system was evaluated for paralleled next-generation sequencing ( NGS) of AGXT, GRHPR, and HOGA1 in 90 known PH patients. AGXT sequencing was requested in all patients, permitting a diagnosis of PH1 in 50%. All remaining patients underwent targeted exon sequencing of GRHPR and HOGA1 with 8% diagnosed with PH2 and 8% with PH3. Complete sequencing of both GRHPR and HOGA1 was not requested in 25% of patients referred leaving their diagnosis in doubt. NGS analysis showed 98% agreement with Sanger sequencing and both approaches had 100% diagnostic specificity. Diagnostic sensitivity of Sanger sequencing was 98% and for NGS it was 97%. NGS has comparable diagnostic performance to Sanger sequencing for the diagnosis of PH and, if implemented, would screen for all forms of PH simultaneously ensuring prompt diagnosis at decreased cost.
- Subjects
MOLECULAR genetics; MOLECULAR biology; DIAGNOSIS; CLINICAL medicine; MEDICAL care
- Publication
Molecular Genetics & Genomic Medicine, 2015, Vol 3, Issue 1, p69
- ISSN
2324-9269
- Publication type
Article
- DOI
10.1002/mgg3.118