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

Lookup NU author(s): Dr Peter Carey



This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


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.

Publication metadata

Author(s): Roy NBA, Wilson EA, Henderson S, Wray K, Babbs C, Okoli S, Atoyebi W, Mixon A, Cahill MR, Carey P, Cullis J, Curtin J, Dreau H, Ferguson DJP, Gibson B, Hall G, Mason J, Morgan M, Proven M, Qureshi A, Garcia JS, Sirachainan N, Teo J, Tedgard U, Higgs D, Roberts D, Roberts I, Schuh A

Publication type: Article

Publication status: Published

Journal: British Journal of Haematology

Year: 2016

Volume: 175

Issue: 2

Pages: 318-330

Print publication date: 01/10/2016

Online publication date: 19/07/2016

Acceptance date: 08/05/2016

Date deposited: 16/01/2017

ISSN (print): 0007-1048

ISSN (electronic): 1365-2141

Publisher: Wiley-Blackwell


DOI: 10.1111/bjh.14221


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Funder referenceFunder name
BRC Blood Theme
BRC/NHS Translational Molecular Diagnostics Centre, Oxford University Hospitals
MRC Molecular Haematology Unit, University of Oxford
National Institute for Health Research (NIHR) Oxford BRC
GN2300Henry Smith Charity and Action Medical Research