Gene diagnostic method for primary immunodeficiency disease based on targeted panel sequencing
10.3969/j.issn.1674-8115.2017.03.022
- VernacularTitle:基于靶向基因测序的原发性免疫缺陷病基因诊断方法
- Author:
Lijun YANG
;
Niu LI
;
Yi LIU
;
Jian WANG
- Keywords:
primary immunodeficiency disease;
targeted panel sequencing;
WHIM syndrome;
CXCR4 gene;
de novo
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2017;37(3):390-393
- CountryChina
- Language:Chinese
-
Abstract:
Objective · To design and build a high-throughput sequencing approach based on targeted panel sequencing (TPS) using for the primary immunodeficiency disease (PID) diagnosis. Methods · By reviewing the literature and querying the relevant databases to determine the known diseasecausing genes of PID, capture probes using for the TPS were designed and customized for all exons and flanking sequences of these genes. A child suspected with PID was diagnosed by the customized TPS. Results · The PID sequencing panel contains a total of 100 known pathogenic genes. The sequencing data of the patient has 16414298 reads. The average coverage depth is 157 X, 98.35% of the target region sequencing depth is greater than 20 X, and 99.97% of the target region sequencing depth is greater than 1 X. Finally, a heterozygous nonsense mutation was found in the exon 2 of the CXCR4 gene (c.1000C>T, p.Arg334*) in the child. The results of Sanger sequencing confirmed the variation in the child and showed that his parents were wildtype at the corresponding sites, indicating the mutation is de novo. Conclusion · This study established a high-throughput sequencing diagnostic approach for PID, with which a case of WHIM syndrome was successfully diagnosed.
