文献_PCR-Based Methods for the Enrichment of Minority Alleles and Mutations.pdf

Clinical Chemistry 55:4 Mini-Reviews 632–640 (2009) PCR-Based Methods for the Enrichment of Minority Alleles and Mutations Coren A. Milbury,1 Jin Li,1 and G. Mike Makrigiorgos1* BACKGROUND: The ability to identify low-level somatic A prominent concern confronting clinical and diag- DNA mutations and minority alleles within an excess nostic applications is the ability to detect clinically sig- wild-type sample is becoming essential for characteriz- nificant low-level mutations and minority alleles. The ing early and posttreatment tumor status in cancer pa- ability to discern mutations is important in many re- tients. Over the past 2 decades, much research has fo- gards, but especially for (a) early cancer detection from cused on improving the selectivity of PCR-based tissue biopsies and bodily fluids such as plasma or se- technologies for enhancing the detection of minority rum; (b) assessment of residual disease after surgery or (mutant) alleles in clinical samples. Routine applica- radiochemotherapy; (c) disease staging and molecular tion in clinical and diagnostic settings requires that profiling for prognosis or tailoring therapy to individ- these techniques be accurate and cost-effective and re- ual patients; and (d) monitoring of therapy outcome quire little effort to optimize, perform, and analyze. and cancer remission/relapse. Efficient detection of cancer-relevant somatic mutations largely depends on CONTENT: Enrichment methods typically segregate by the selectivity of the techniques and methods em- their abi