TY - JOUR
T1 - Rapid molecular diagnostics of severe primary immunodeficiency determined by using targeted next-generation sequencing
AU - Yu, Hui
AU - Zhang, Victor Wei
AU - Stray-Pedersen, Asbjørg
AU - Hanson, Imelda Celine
AU - Forbes, Lisa R.
AU - de la Morena, M. Teresa
AU - Chinn, Ivan K.
AU - Gorman, Elizabeth
AU - Mendelsohn, Nancy J.
AU - Pozos, Tamara
AU - Wiszniewski, Wojciech
AU - Nicholas, Sarah K.
AU - Yates, Anne B.
AU - Moore, Lindsey E.
AU - Berge, Knut Erik
AU - Sorte, Hanne
AU - Bayer, Diana K.
AU - ALZahrani, Daifulah
AU - Geha, Raif S.
AU - Feng, Yanming
AU - Wang, Guoli
AU - Orange, Jordan S.
AU - Lupski, James R.
AU - Wang, Jing
AU - Wong, Lee Jun
N1 - Publisher Copyright:
© 2016 American Academy of Allergy, Asthma & Immunology
PY - 2016/10/1
Y1 - 2016/10/1
N2 - Background Primary immunodeficiency diseases (PIDDs) are inherited disorders of the immune system. The most severe form, severe combined immunodeficiency (SCID), presents with profound deficiencies of T cells, B cells, or both at birth. If not treated promptly, affected patients usually do not live beyond infancy because of infections. Genetic heterogeneity of SCID frequently delays the diagnosis; a specific diagnosis is crucial for life-saving treatment and optimal management. Objective We developed a next-generation sequencing (NGS)–based multigene-targeted panel for SCID and other severe PIDDs requiring rapid therapeutic actions in a clinical laboratory setting. Methods The target gene capture/NGS assay provides an average read depth of approximately 1000×. The deep coverage facilitates simultaneous detection of single nucleotide variants and exonic copy number variants in one comprehensive assessment. Exons with insufficient coverage (<20× read depth) or high sequence homology (pseudogenes) are complemented by amplicon-based sequencing with specific primers to ensure 100% coverage of all targeted regions. Results Analysis of 20 patient samples with low T-cell receptor excision circle numbers on newborn screening or a positive family history or clinical suspicion of SCID or other severe PIDD identified deleterious mutations in 14 of them. Identified pathogenic variants included both single nucleotide variants and exonic copy number variants, such as hemizygous nonsense, frameshift, and missense changes in IL2RG; compound heterozygous changes in ATM, RAG1, and CIITA; homozygous changes in DCLRE1C and IL7R; and a heterozygous nonsense mutation in CHD7. Conclusion High-throughput deep sequencing analysis with complete clinical validation greatly increases the diagnostic yield of severe primary immunodeficiency. Establishing a molecular diagnosis enables early immune reconstitution through prompt therapeutic intervention and guides management for improved long-term quality of life.
AB - Background Primary immunodeficiency diseases (PIDDs) are inherited disorders of the immune system. The most severe form, severe combined immunodeficiency (SCID), presents with profound deficiencies of T cells, B cells, or both at birth. If not treated promptly, affected patients usually do not live beyond infancy because of infections. Genetic heterogeneity of SCID frequently delays the diagnosis; a specific diagnosis is crucial for life-saving treatment and optimal management. Objective We developed a next-generation sequencing (NGS)–based multigene-targeted panel for SCID and other severe PIDDs requiring rapid therapeutic actions in a clinical laboratory setting. Methods The target gene capture/NGS assay provides an average read depth of approximately 1000×. The deep coverage facilitates simultaneous detection of single nucleotide variants and exonic copy number variants in one comprehensive assessment. Exons with insufficient coverage (<20× read depth) or high sequence homology (pseudogenes) are complemented by amplicon-based sequencing with specific primers to ensure 100% coverage of all targeted regions. Results Analysis of 20 patient samples with low T-cell receptor excision circle numbers on newborn screening or a positive family history or clinical suspicion of SCID or other severe PIDD identified deleterious mutations in 14 of them. Identified pathogenic variants included both single nucleotide variants and exonic copy number variants, such as hemizygous nonsense, frameshift, and missense changes in IL2RG; compound heterozygous changes in ATM, RAG1, and CIITA; homozygous changes in DCLRE1C and IL7R; and a heterozygous nonsense mutation in CHD7. Conclusion High-throughput deep sequencing analysis with complete clinical validation greatly increases the diagnostic yield of severe primary immunodeficiency. Establishing a molecular diagnosis enables early immune reconstitution through prompt therapeutic intervention and guides management for improved long-term quality of life.
KW - Severe combined immunodeficiency
KW - molecular diagnostics
KW - next-generation sequencing
KW - severe combined immunodeficiency newborn screening
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U2 - 10.1016/j.jaci.2016.05.035
DO - 10.1016/j.jaci.2016.05.035
M3 - Article
C2 - 27484032
AN - SCOPUS:84992170316
SN - 0091-6749
VL - 138
SP - 1142-1151.e2
JO - Journal of Allergy and Clinical Immunology
JF - Journal of Allergy and Clinical Immunology
IS - 4
ER -