A novel 33-Gene targeted resequencing panel provides accurate, clinical-grade diagnosis and improves patient management for rare inherited anaemias

作者：Roy Noemi B A; Wilson Edward A; Henderson Shirley; Wray Katherine; Babbs Christian; Okoli Steven; Atoyebi Wale; Mixon Avery; Cahill Mary R; Carey Peter; Cullis Jonathan; Curtin Julie; Dreau Helene; Ferguson David J P; Gibson Brenda; Hall Georgina; Mason Joanne; Morgan Mary; Proven Melanie; Qureshi Amrana; Sanchez Garcia Joaquin; Sirachainan Nongnuch; Teo Juliana; Tedgard Ulf; Higgs Doug; Roberts David; Roberts Irene^*; Schuh Anna^*

来源：British Journal of Haematology, 2016, 175(2): 318-330.

DOI：10.1111/bjh.14221

• inherited anaemia
• congenital dyserythropoietic anaemia
• molecular genetics
• pyruvate kinase deficiency
• next-generation sequencing

Accurate diagnosis of rare inherited anaemias is challenging, requiring a series of complex and expensive laboratory tests. Targeted next-generation-sequencing (NGS) has been used to investigate these disorders, but the selection of genes on individual panels has been narrow and the validation strategies used have fallen short of the standards required for clinical use. Clinical-grade validation of negative results requires the test to distinguish between lack of adequate sequencing reads at the locations of known mutations and a real absence of mutations. To achieve a clinically-reliable diagnostic test and minimize false-negative results we developed an open-source tool (CoverMi) to accurately determine base-coverage and the discoverability' of known mutations for every sample. We validated our 33-gene panel using Sanger sequencing and microarray. Our panel demonstrated 100% specificity and 997% sensitivity. We then analysed 57 clinical samples: molecular diagnoses were made in 22/57 (386%), corresponding to 32 mutations of which 16 were new. In all cases, accurate molecular diagnosis had a positive impact on clinical management. Using a validated NGS-based platform for routine molecular diagnosis of previously undiagnosed congenital anaemias is feasible in a clinical diagnostic setting, improves precise diagnosis and enhances management and counselling of the patient and their family.

• 出版日期2016-10