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목적 : To establish initial set-up of next-generation sequencing (NGS) technology in university-based ophthalmology practice including neuro-ophthalmology and find responsible genes for ophthalmologic genetic disorders.
방법 : Target gene library was construct to capture exons of responsible genes using hybridization capture technique. Each patient’s genomic DNA was fragmented with a median size of 150 base pair. Pooled libraries were sequence on a NextSeq 550 (Illumina, San Diego, CA) using the NextSeq 500/550 v2 Reagent kit.
결과 : PubMed searches using term NGS, congenital cataract, leber congenital amaurosis, inherited retinal dystrophy, retinitis pigmentosa and assessment of RetNet were done to identify list of genes responsible for eye disorders. Gene list in other laboratories including MEEI, Emory Eye Center, CEN4GEN, and Fulgent were searched to identify any missing genes. All known intronic variant were retrieved by human gene mutation database and literature searches. A total 411 genes were included in the Pan-eye panel of Severance Hospital.
결론 : Target 411 genes including retinal dystrophy, infantile nystagmus syndrome, Leber congenital amaurosis will aid accurate molecular diagnosis in ophthalmologic practice. Although NGS is not a magic bullet and tremendous effort is needed to process bioinformatics analysis, NGS gained more popularity because phenotypes of these disorders are similar in infant with poor vision and cost of sequencing is decreasing continuously compared to Sanger sequencing.
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