Diagnosis of Noonan syndrome and related disorders using target next generation sequencing

BMC Medical Genetics

Bruno Dallapiccola²  Adriano Angioni²  Tiziana Franchin²  Serena Russo²  Anwar Baban²  Rossella Capolino²  Pietro Sirleto²  Elisa Pisaneschi²  Maria Lisa Dentici²  Simona Grotta²  Maria Gnazzo²  Maria Cristina Digilio²  Rossana Scavelli¹  Francesca Romana Lepri² 

[1] Illumina, Inc., San Diego, CA 92122, USA;Cytogenetics, Medical Genetics and Pediatric Cardiology, Bambino Gesù Children Hospital, IRCCS, Rome, Italy

关键词: RASopathies;    Molecular diagnosis;    Next generation sequencing;    Noonan syndrome;   

Others  :  1122522
DOI  :  10.1186/1471-2350-15-14

 received in 2013-09-25, accepted in 2014-01-20,  发布年份 2014

【 摘 要 】

Background

Noonan syndrome is an autosomal dominant developmental disorder with a high phenotypic variability, which shares clinical features with other rare conditions, including LEOPARD syndrome, cardiofaciocutaneous syndrome, Noonan-like syndrome with loose anagen hair, and Costello syndrome. This group of related disorders, so-called RASopathies, is caused by germline mutations in distinct genes encoding for components of the RAS-MAPK signalling pathway. Due to high number of genes associated with these disorders, standard diagnostic testing requires expensive and time consuming approaches using Sanger sequencing. In this study we show how targeted Next Generation Sequencing (NGS) technique can enable accurate, faster and cost-effective diagnosis of RASopathies.

Methods

In this study we used a validation set of 10 patients (6 positive controls previously characterized by Sanger-sequencing and 4 negative controls) to assess the analytical sensitivity and specificity of the targeted NGS. As second step, a training set of 80 enrolled patients with a clinical suspect of RASopathies has been tested. Targeted NGS has been successfully applied over 92% of the regions of interest, including exons for the following genes: PTPN11, SOS1, RAF1, BRAF, HRAS, KRAS, NRAS, SHOC, MAP2K1, MAP2K2, CBL.

Results

All expected variants in patients belonging to the validation set have been identified by targeted NGS providing a detection rate of 100%. Furthermore, all the newly detected mutations in patients from the training set have been confirmed by Sanger sequencing. Absence of any false negative event has been excluded by testing some of the negative patients, randomly selected, with Sanger sequencing.

Conclusion

Here we show how molecular testing of RASopathies by targeted NGS could allow an early and accurate diagnosis for all enrolled patients, enabling a prompt diagnosis especially for those patients with mild, non-specific or atypical features, in whom the detection of the causative mutation usually requires prolonged diagnostic timings when using standard routine. This approach strongly improved genetic counselling and clinical management.

【 授权许可】

   
2014 Lepri et al.; licensee BioMed Central Ltd.

【 图 表 】

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