Next-generation sequencing in childhood disorders

Abstract

Genetics has been revolutionised by recent technologies. The latest addition to these advances is next-generation sequencing, which is set to transform clinical diagnostics in every branch of medicine. In the research arena this has already been instrumental in identifying hundreds of novel genetic syndromes, making a molecular diagnosis possible for the first time in numerous refractory cases. However, the pace of change has left many clinicians bewildered by new terminology and the implications of next-generation sequencing for their clinical practice. The rapid developments have also left many diagnostic laboratories struggling to implement these new technologies with limited resources. This review explains the basic concepts of next-generation sequencing, gives examples of its role in clinically applied research and examines the challenges of its introduction into clinical practice.

Candidate gene discovery in autoimmunity by using extreme phenotypes, next generation sequencing and whole exome capture

“…The formed ends are adenylated and adaptor oligonucleotides are added to the ends of these adenylated fragments. These adaptors are short oligonucleotides of known sequences for universal priming of both amplification and sequencing steps [5]. Commonly, the fragments are enriched for specific genes of interest (targeted sequencing), or for all coding regions (whole exome capture for WES), in a physical capture step.…”

“…Commonly, the fragments are enriched for specific genes of interest (targeted sequencing), or for all coding regions (whole exome capture for WES), in a physical capture step. That enrichment is not needed for WGS and all fragments are sequenced [5].…”

“…Whole exome sequencing (WES) is a cost effective technique that employs high throughput capture by hybridization techniques, using exon specific oligonucleotides to enrich only protein coding sequences that can be later used for sequencing [5]. The WES is rapidly becoming the first-line approach for monogenic disorders, and an alternative one for dissecting extreme phenotypes of complex inherited conditions [4,6,7].…”

Next-generation sequencing identifies articular cartilage and subchondral bone miRNAs after ESWT on early osteoarthritis knee

“…Several miRNAs are demonstrated that associated with OA development and modulation such as miR-18a (chondrocyte differentiation), miR-27b (controlling the expression of MMP-13), miR-34a (prevention of cartilage degradation), miR-140 and miR-222 (controlling cartilage homeostasis), miR-146 (promotion of inflammatory OA), miR146a (OA cartilage pathogenesis), miR-675 (cartilage repair) [30, 31]. With the development of NGS technology, changes in the expression levels of thousands of genes and miRNAs can be examined simultaneously, and integral analysis of the dysregulated genes can be performed to obtain information regarding pathogenic mechanisms of OA at the cellular level, animal model and human disease [32, 33]. Furthermore, NGS data can be used to discover novel molecular diagnostic markers and therapeutic targets [34].…”

Genomics in early infantile epileptic encephalopathies – trials and tribulations

“…In the latter case this is often because of a lack of identified candidate genes. In these cases, exomic/genomic sequencing of panels of known genes causing EIEEs will identify a considerable number . Although more expensive than some investigations, the cost benefit is likely to be dramatically better than previous technology.…”