Integration of Next-generation sequencingtechnologies in pathological diagnostics

Abstract

Today, with the better understanding of the molecular events involved in malignancy and the mechanisms of pharmacotherapy, larger gene panels are more helpful than single biomarker detection. After the completion of the first human genome sequence in 2004, the growing need to sequence a large number of individual genomes in a fast, low-cost and accurate way has directed a shift from traditional Sanger sequencing methods towards new high-throughput genomic technologies. In 2005, the development of next generation sequencing (NGS) methods has represented one of the more significant technical advances in molecular biology. NGS, also known as massive parallel sequencing because of the ability to allow the parallel analysis of a very large number of DNA molecules, is beginning to show its full potential for diagnostic and therapeutic applications. Until recently, NGS platforms were envisioned for large-scale applications,focused on whole genome sequencing, with protocols, consumable costs and a turnaround time (TAT) unsuitable for the needs of small diagnostic laboratories. The development of miniaturised technology by benchtop NGS sequencers decreased sequencing costs, moving NGS from a few large sequencing core centers to a much larger number of individual laboratories. Currently, most pathology departments acquired and NGS benchtop sequencer, thus NGS is adopted for routine molecular diagnostics,including cytological samples. To understand the current and future application of NGS in the field of pathology,modern pathologists need to understand its basic principles. This thesis describes my research on the integration of NGS technologies in pathological diagnostics, both concerning histological and cytological specimens. Moreover, a research application of NGS on mouse xenograft cytological samples is described.