RT Journal Article
T1 A novel 33-Gene targeted resequencing panel provides accurate, clinical-grade diagnosis and improves patient management for rare inherited anaemias.
A1 Roy, Noémi B A
A1 Wilson, Edward A
A1 Henderson, Shirley
A1 Wray, Katherine
A1 Babbs, Christian
A1 Okoli, Steven
A1 Atoyebi, Wale
A1 Mixon, Avery
A1 Cahill, Mary R
A1 Carey, Peter
A1 Cullis, Jonathan
A1 Curtin, Julie
A1 Dreau, Helene
A1 Ferguson, David J P
A1 Gibson, Brenda
A1 Hall, Georgina
A1 Mason, Joanne
A1 Morgan, Mary
A1 Proven, Melanie
A1 Qureshi, Amrana
A1 Sanchez Garcia, Joaquin
A1 Sirachainan, Nongnuch
A1 Teo, Juliana
A1 Tedgård, Ulf
A1 Higgs, Doug
A1 Roberts, David
A1 Roberts, Irene
A1 Schuh, Anna
K1 congenital dyserythropoietic anaemia
K1 inherited anaemia
K1 molecular genetics
K1 next-generation sequencing
K1 pyruvate kinase deficiency
AB 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 99·7% sensitivity. We then analysed 57 clinical samples: molecular diagnoses were made in 22/57 (38·6%), 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.
YR 2016
FD 2016-07-19
LK http://hdl.handle.net/10668/10282
UL http://hdl.handle.net/10668/10282
LA en
DS RISalud
RD Apr 17, 2025