- 作者：Isaac J. Nijman ; Joris M. van Montfrans ; Marlous Hoogstraat ; Marianne L. Boes ; Lisette van de Corput ; Ellen D. Renner ; Patrick van Zon ; Stef van Lieshout ; Martin G. Elferink ; Mirjam van der Burg ; Clementien L. Vermont ; Bert van der Zwaag ; Esther Janson ; Edwin Cuppen ; Johannes K. Ploos van Amstel ; Marielle E. van Gijn
- 关键词：Primary immunodeficiency ; diagnosis ; genetics ; next-generation sequencing
- 刊名：Journal of Allergy and Clinical Immunology
- 出版年：February, 2014
- 年：2014
- 卷：133
- 期：2
- 页码：529-534.e1
- 全文大小：188 K
ss="h4">Background
Primary immunodeficiency (PID) disorders are a heterogeneous group of inherited disorders caused by a variety of聽monogenetic immune defects. Thus far, mutations in more than 170聽different genes causing PIDs have been described. A聽clear genotype-phenotype correlation is often not available, which makes a genetic diagnosis in patients with PIDs complex and laborious.ss="h4">Objective
We sought to develop a robust, time-effective, and cost-effective diagnostic method to facilitate a genetic diagnosis in any of 170 known PID-related genes by using next-generation sequencing (NGS).
ss="h4">Methods
We used both targeted array-based and in-solution enrichment combined with a SOLiD sequencing platform and a聽bioinformatic pipeline developed in house to analyze genetic changes in the DNA of 41 patients with PIDs with known mutations and 26 patients with undiagnosed PIDs.
ss="h4">Results
This novel NGS-based method accurately detected point mutations (sensitivity and specificity >99% in covered regions) and exonic deletions (100% sensitivity and specificity). For the 170 genes of interest, the DNA coverage was greater than 20脳 in 90% to 95%. Nine PID-related genes proved not eligible for evaluation by using this NGS-based method because of inadequate coverage. The NGS method allowed us to make a genetic diagnosis in 4 of 26 patients who lacked a genetic diagnosis despite routine functional and genetic testing. Three of聽these patients proved to have an atypical presentation of previously described PIDs.
ss="h4">Conclusion
This novel NGS tool facilitates accurate simultaneous detection of mutations in 161 of 170 known PID-related genes. In addition, these analyses will generate more insight into genotype-phenotype correlations for the different PID disorders.