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

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A novel 33-Gene targeted resequencing panel provides accurate, clinical-grade diagnosis and improves patient management for rare inherited anaemias. / Roy, Noémi 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; Tedgård, Ulf; Higgs, Doug; Roberts, David; Roberts, Irene; Schuh, Anna.

In: British Journal of Haematology, Vol. 175, No. 2, 10.2016, p. 318-330.

Research output: Contribution to journalArticle

Harvard

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, Sanchez Garcia, J, Sirachainan, N, Teo, J, Tedgård, U, Higgs, D, Roberts, D, Roberts, I & Schuh, A 2016, 'A novel 33-Gene targeted resequencing panel provides accurate, clinical-grade diagnosis and improves patient management for rare inherited anaemias', British Journal of Haematology, vol. 175, no. 2, pp. 318-330. https://doi.org/10.1111/bjh.14221

APA

CBE

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, Sanchez Garcia J, Sirachainan N, Teo J, Tedgård U, Higgs D, Roberts D, Roberts I, Schuh A. 2016. A novel 33-Gene targeted resequencing panel provides accurate, clinical-grade diagnosis and improves patient management for rare inherited anaemias. British Journal of Haematology. 175(2):318-330. https://doi.org/10.1111/bjh.14221

MLA

Vancouver

Author

Roy, Noémi 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 ; Tedgård, Ulf ; Higgs, Doug ; Roberts, David ; Roberts, Irene ; Schuh, Anna. / A novel 33-Gene targeted resequencing panel provides accurate, clinical-grade diagnosis and improves patient management for rare inherited anaemias. In: British Journal of Haematology. 2016 ; Vol. 175, No. 2. pp. 318-330.

RIS

TY - JOUR

T1 - A novel 33-Gene targeted resequencing panel provides accurate, clinical-grade diagnosis and improves patient management for rare inherited anaemias

AU - Roy, Noémi B A

AU - Wilson, Edward A.

AU - Henderson, Shirley

AU - Wray, Katherine

AU - Babbs, Christian

AU - Okoli, Steven

AU - Atoyebi, Wale

AU - Mixon, Avery

AU - Cahill, Mary R.

AU - Carey, Peter

AU - Cullis, Jonathan

AU - Curtin, Julie

AU - Dreau, Helene

AU - Ferguson, David J P

AU - Gibson, Brenda

AU - Hall, Georgina

AU - Mason, Joanne

AU - Morgan, Mary

AU - Proven, Melanie

AU - Qureshi, Amrana

AU - Sanchez Garcia, Joaquin

AU - Sirachainan, Nongnuch

AU - Teo, Juliana

AU - Tedgård, Ulf

AU - Higgs, Doug

AU - Roberts, David

AU - Roberts, Irene

AU - Schuh, Anna

PY - 2016/10

Y1 - 2016/10

N2 - 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.

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.

KW - Congenital dyserythropoietic anaemia

KW - Inherited anaemia

KW - Molecular genetics

KW - Next-generation sequencing

KW - Pyruvate kinase deficiency

UR - http://www.scopus.com/inward/record.url?scp=84978634344&partnerID=8YFLogxK

U2 - 10.1111/bjh.14221

DO - 10.1111/bjh.14221

M3 - Article

VL - 175

SP - 318

EP - 330

JO - British Journal of Haematology

T2 - British Journal of Haematology

JF - British Journal of Haematology

SN - 0007-1048

IS - 2

ER -