Performance evaluation of Sanger sequencing for the diagnosis of primary hyperoxaluria and comparison with targeted next generation sequencing. Issue 1 (5th November 2014)

• Record Type: Journal Article
• Authors: Williams, Emma L.; Bagg, Eleanor A. L.; Mueller, Michael; Vandrovcova, Jana; Aitman, Timothy J.; Rumsby, Gill
• Is Part Of: Molecular genetics & genomic medicine. Volume 3:Issue 1(2015:Jan.)
• Page Start: 69
• Page End: 78
• Publication Date: 2014-11-05
• Subjects: Medical genetics -- Periodicals; Genomics -- Periodicals; 616.042
• DOI: 10.1002/mgg3.118
• Languages: English
• ISSNs: 2324-9269

Abstract

<abstract abstract-type="main" id="mgg3118-abs-0001"> <title>Abstract</title> <p>Definitive diagnosis of primary hyperoxaluria (PH) currently utilizes sequential Sanger sequencing of the <italic>AGXT, GRPHR</italic>, and <italic>HOGA1</italic> 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 <italic>AGXT</italic>, <italic> GRHPR</italic>, and <italic>HOGA1</italic> in 90 known PH patients. <italic>AGXT</italic> sequencing was requested in all patients, permitting a diagnosis of PH1 in 50%. All remaining patients underwent targeted exon sequencing of <italic>GRHPR</italic> and <italic>HOGA1</italic> with 8% diagnosed with PH2 and 8% with PH3. Complete sequencing of both <italic>GRHPR</italic> and <italic>HOGA1</italic> 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.</p> </abstract>