ABSTRACT
Twenty-’rst century medical practice can be predicted to bene’t from a con™uence of scienti’c and technologic advances leading toward truly rational and personalized health care. These advances include thorough understanding of the human genome including sequence-function relationships, the role of regulatory regions, and the effects of genetic variation on phenotypic expression; comprehensive structural modeling of DNA, ribonucleic acids, proteins, small molecules, and their speci’c interactions so that rational drug design can be perfected; systems biology approaches to understanding the complex networks of metabolic, signal transduction, and regulatory pathways at the organelle, cellular, organ, and organism levels so that targeted therapies can be optimized with minimal side effects; and exponential growth in computer hardware and software to process the increasing complexity of the above developments. A particular aspect of personalized medicine is the study of the effects of genetic variation in the response to drug therapy, which can be labeled as pharmacogenomics when the focus is on largescale analyses of the genome, or pharmacogenetics, when the focus is on a relatively restricted number of genes. This distinction is becoming fuzzier as newer technologies for genetic analysis exhibit higher multiplexing abilities and lower cost per assay and if whole genomic testing becomes routine and replaces focused genetic testing. This chapter brie™y describes a variety of methods used to identify DNA variation affecting genes involved in drug metabolism (pharmacokinetics) or
10.1 Introduction .................................................................................................. 211 10.2 Low Multiplexing Assays ............................................................................. 212 10.3 Mid-Multiplexing Assays ............................................................................. 219 10.4 High Multiplexing and Next-Generation Sequencing Systems .................... 222
10.4.1 High-Plex Single Nucleotide Polymorphism (SNP) Assays ............. 222 10.4.2 High-Throughput Library Preparation and Polymerase Chain
Reaction (PCR) Systems ................................................................... 222 10.4.3 Next-Generation Sequencing ............................................................224
10.5 Conclusions ................................................................................................... 227 References .............................................................................................................. 227
drug target genes (pharmacodynamics), starting from currently available methods focused on a restricted set of genes and ending on high-throughput next-generation sequencing. Despite the rapid development of technologic advances and the multitude of commercially available research-use-only reagents for pharmacogenetic testing, in June 2011 only nine pharmacogenetics assays were U.S. Food and Drug Administration (FDA)-approved, of which ’ve are used for Warfarin PGx (Table 10.1). It is important that the legal and regulatory environment keeps up with the rapid pace of development in genetics to fully realize its bene’ts to human health.
