The Application Of Next-generation Sequencing And Exon Trapping Technology In Genetic Molecular Diagnosis Of Genetic Disorders

There are multiple molecular mechanisms that can cause genetic disorders such as point mutations,splicing site mutation, insertion/deletion mutations, gene duplication, complex rearrangements and gene copy number abnormalities. As an important analysis method of molecular biology, DNA sequencing technology has been widely used in gene detection and disease diagnosis, etc. With DNA sequencing technology, we can detect most point mutation and some small insertions/deletions mutations. However, due to the large size of many genes and genetic heterogeneity of many diseases, the Sanger sequencing technology has been far from satisfying the needs of scientific research and clinical applications. The next-generation sequencing technology has emerged at the right time. Due to the lower cost and much higher throughput, it has been widely used in scientific research and clinical practics.Exon trapping technology is another useful tool in genetic diseases diagnosis. For those point mutations which may cause splice site changes, we can use this technology to obtain an accurate diagnosis and insight of the pathogenic mechanisms. Although there are a variety of prediction software can be used to predict the splice site changes, but these predictions still need experimental verification. So when we undertake a clinical diagnosis, both exon trapping technology and predictive analytics are needed to get an accurate result.In this study, we use the next-generation sequencing technology and exon trapping technology in three clinical cases of16p11.2microdeletion syndrome, Leber’s congenital amaurosis and X-linked Hypohidrotic Ectodermal Dysplasia. Our data confirmed that these two techniques could be widely used in clinical diagnosis of genetic disorders. But we also identified some limitations for these two techniques. Multiple methods should be used for correct diagnosis.