Whole exome sequencing is an efficient and sensitive method for detection of germline mutations in patients with phaeochromcytomas and paragangliomas. (25th October 2013)

• Record Type: Journal Article
• Authors: McInerney‐Leo, Aideen M.; Marshall, Mhairi S.; Gardiner, Brooke; Benn, Diana E.; McFarlane, Janelle; Robinson, Bruce G.; Brown, Matthew A.; Leo, Paul J.; Clifton‐Bligh, Roderick J.; Duncan, Emma L.
• Is Part Of: Clinical endocrinology. Volume 80:Number 1(2014:Jan.)
• Page Start: 25
• Page End: 33
• Publication Date: 2013-10-25
• Subjects: Endocrinology -- Periodicals; 616.4005
• DOI: 10.1111/cen.12331
• Languages: English
• ISSNs: 0300-0664

Abstract

<abstract abstract-type="main" id="cen12331-abs-0001"> <title>Summary</title> <sec id="cen12331-sec-0001" sec-type="section"> <title>Background</title> <p>Genetic testing is recommended when the probability of a disease‐associated germline mutation exceeds 10%. Germline mutations are found in approximately 25% of individuals with phaeochromcytoma (PCC) or paraganglioma (PGL); however, genetic heterogeneity for PCC/PGL means many genes may require sequencing. A phenotype‐directed iterative approach may limit costs but may also delay diagnosis, and will not detect mutations in genes not previously associated with PCC/PGL.</p> </sec> <sec id="cen12331-sec-0002" sec-type="section"> <title>Objective</title> <p>To assess whether whole exome sequencing (WES) was efficient and sensitive for mutation detection in PCC/PGL.</p> </sec> <sec id="cen12331-sec-0003" sec-type="section"> <title>Methods</title> <p>Whole exome sequencing was performed on blinded samples from eleven individuals with PCC/PGL and known mutations. Illumina TruSeq™ (Illumina Inc, San Diego, CA, USA) was used for exome capture of seven samples, and NimbleGen SeqCap EZ v3.0 (Roche NimbleGen Inc, Basel, Switzerland) for five samples (one sample was repeated). Massive parallel sequencing was performed on multiplexed samples. Sequencing data were called using Genome Analysis Toolkit and annotated using <sc>annovar</sc>. Data were assessed for coding variants in <italic>RET, NF1, VHL, SDHD, SDHB, SDHC, SDHA, SDHAF2, KIF1B, TMEM127, EGLN1</italic> and <italic>MAX</italic>. Target capture of five exome capture platforms was compared.</p> </sec> <sec id="cen12331-sec-0004" sec-type="section"> <title>Results</title> <p>Six of seven mutations were detected using Illumina TruSeq™ exome capture. All five mutations were detected using NimbleGen SeqCap EZ v3.0 platform, including the mutation missed using Illumina TruSeq™ capture. Target capture for exons in known PCC/PGL genes differs substantially between platforms. Exome sequencing was inexpensive (<$A800 per sample for reagents) and rapid (results <5 weeks from sample reception).</p> </sec> <sec id="cen12331-sec-0005" sec-type="section"> <title>Conclusion</title> <p>Whole exome sequencing is sensitive, rapid and efficient for detection of PCC/PGL germline mutations. However, capture platform selection is critical to maximize sensitivity.</p> </sec> </abstract>