Rapid molecular diagnostics of severe primary immunodeficiency determined by using targeted next-generation sequencing

被引：59

作者：

Yu, Hui ^{[1
]}

Zhang, Victor Wei ^{[2
]}

Stray-Pedersen, Asbjorg ^{[3
,4
,5
,6
,7
]}

Hanson, Imelda Celine ^{[3
,4
,5
]}

Forbes, Lisa R. ^{[3
,4
,5
]}

de la Morena, M. Teresa ^{[9
]}

Chinn, Ivan K. ^{[3
,4
,5
]}

Gorman, Elizabeth ^{[1
]}

Mendelsohn, Nancy J. ^{[10
]}

Pozos, Tamara ^{[11
]}

Wiszniewski, Wojciech ^{[6
]}

Nicholas, Sarah K. ^{[3
,4
,5
]}

Yates, Anne B. ^{[12
]}

Moore, Lindsey E. ^{[12
]}

Berge, Knut Erik ^{[8
]}

Sorte, Hanne ^{[8
]}

Bayer, Diana K. ^{[13
]}

ALZahrani, Daifulah ^{[14
]}

Geha, Raif S. ^{[15
,16
]}

Feng, Yanming ^{[1
]}

Wang, Guoli ^{[1
]}

Orange, Jordan S. ^{[3
,4
,5
]}

Lupski, James R. ^{[2
,4
,6
,17
]}

Wang, Jing ^{[2
]}

Wong, Lee-Jun ^{[2
]}

机构：

[1] Baylor Genet, Houston, TX USA

[2] Baylor Coll Med, Dept Mol & Human Genet, One Baylor Plaza,NAB 2015, Houston, TX 77030 USA

[3] Baylor Coll Med, Dept Pediat, Sect Immunol Allergy & Rheumatol, Houston, TX 77030 USA

[4] Texas Childrens Hosp, Houston, TX 77030 USA

[5] Texas Childrens Hosp, Feigin Ctr, Ctr Human Immunobiol Immunol Allergy & Rheumatol, Houston, TX 77030 USA

[6] Baylor Coll Med, Baylor Hopkins Ctr Mendelian Genom, Dept Mol & Human Genet, Houston, TX 77030 USA

[7] Oslo Univ Hosp, Norwegian Natl Unit Newborn Screening, Oslo, Norway

[8] Oslo Univ Hosp, Dept Med Genet, Oslo, Norway

[9] Univ Texas Southwestern Med Ctr Dallas, Div Allergy & Immunol, Childrens Med Ctr, Dallas, TX 75390 USA

[10] Childrens Hosp & Clin Minnesota, Minneapolis, MN USA

[11] Childrens Hosp & Clin Minnesota, Pediat Infect Dis & Immunol, St Paul, MN USA

[12] Univ Mississippi, Med Ctr, Jackson, MS 39216 USA

[13] Univ Iowa, Dept Pediat, Div Pediat Allergy Immunol & Pulmonol, Carver Coll Med, Iowa City, IA 52242 USA

[14] King Saud Bin Abdulaziz Univ Hlth Sci, Dept Pediat, Jeddah, Saudi Arabia

[15] Boston Childrens Hosp, Div Immunol, Boston, MA USA

[16] Harvard Med Sch, Dept Pediat, Boston, MA USA

[17] Baylor Coll Med, Dept Pediat, Houston, TX 77030 USA

来源：

JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY | 2016年 / 138卷 / 04期

基金：

美国国家卫生研究院;

关键词：

Severe combined immunodeficiency; severe combined immunodeficiency newborn screening; next-generation sequencing; molecular diagnostics; TBX1; HAPLOINSUFFICIENCY; THYMUS TRANSPLANTATION; OMENN SYNDROME; MUTATIONS; DEFECTS; DISEASE; INFECTIONS; EXPERIENCE; DELETIONS; ARTEMIS;

D O I：

10.1016/j.jaci.2016.05.035

中图分类号：

R392 [医学免疫学];

学科分类号：

100102 ;

摘要：

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 10003. The deep coverage facilitates simultaneous detection of single nucleotide variants and exonic copy number variants in one comprehensive assessment. Exons with insufficient coverage (<20 x 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.

引用

页码：1142 / +

页数：12

共 63 条

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