Abstract 5597: Targeted massively parallel sequencing for the diagnosis of hereditary neuroendocrine disorders

Abstract Background: Genetic testing is a powerful tool for the diagnosis, management and treatment of inherited disease but these tests traditionally have been time-consuming, labour-intensive and costly. Significant improvements in sequencing technologies have the potential to overcome these limitations, through the simultaneous sequencing of multiple genes in multiple samples. Objective: The aim of this study was to develop a multi-gene panel, comprised of hereditary neuroendocrine associated genes, to enable parallel testing of known susceptibility genes; and to determine whether the sensitivity of massively parallel targeted sequencing is equivalent to Sanger sequencing, the current gold standard for diagnostic genetic testing. Methods: We developed a custom gene panel, covering the coding regions of 8 known neuroendocrine suseptibility genes (MAX, SDHB, SDHC, SDHD, SDHAF2, RET, TMEM127 and VHL). In total, genomic DNA of 85 individuals with unique variants in five of the susceptibility genes (based on Sanger sequencing) was prepared using the TruSeq Custom Amplicon Assay (Illumina) and sequenced on an Illumina MiSeq™ platform. Results: Overall, 95.1% of the target was sequenced adequately for variant calling (ie. reads with ≥Q30). Inadequate coverage of the target region was observed for VHL (exon 1), SDHC (exons 2 and 5) and SDHD (exon 4). Seventy-eight of the 85 known variants met the required minimum coverage threshold (7 fell within regions of inadequate coverage or depth of coverage was ≤40). Of these, one variant was undetected, giving an overall sensitivity of 98.7%. Conclusions: Genetic testing using targeted capture followed by massively parallel sequencing, in conjunction with confirmation by Sanger sequencing, is a viable option to the current approach for diagnostic services. Citation Format: Trish Dwight, Anne Louise Richardson, Ying Zhu, Bruce G. Robinson, Diana E. Benn, Roderick J. Clifton-Bligh. Targeted massively parallel sequencing for the diagnosis of hereditary neuroendocrine disorders. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5597. doi:10.1158/1538-7445.AM2014-5597