Next-generation sequencing (NGS) in anatomic pathology discovery and practice

Abstract

Next-generation sequencing (NGS), also known as massively parallel sequencing (MPS), is a family of emerging and powerful technologies that generate an enormous amount of nucleic acid sequence data on a time scale and at a cost that is becoming compatible with routine clinical diagnostic applications. NGS relies upon selective amplification or capture of a complex population of nucleic acid molecules within a biological sample. Simultaneous sequencing reactions generate a variably broad and deep digital readout of sequences in the sample, representing specific regions of genes such as exons, a class of RNAs, or whole genomes. NGS can be viewed as a special type of multianalyte "omics" test that overturns the "one-question/one-test" paradigm prevalent in pathology and laboratory medicine for over a century. It has the potential to address thousands of clinically relevant questions at once while also serving as a transformative, hypothesis-generating discovery tool. The power of NGS to generate massive datasets, which by their sheer size and complexity can approach the level of biological complexity within the tissue sample itself, qualifies it as a "disruptive technology" that will task the medical establishment, particularly pathology and laboratory medicine professionals who generate and interpret the data, to implement it in clinical settings. There remain numerous challenges to the routine implementation of NGS technologies in drug development and clinical diagnostics, including scientific/technical, medical, financial, legal/regulatory, and ethical hurdles.