Enzymatic mutation detection technologies - BioTechniques

REVIEW

Enzymatic mutation detection technologies Anthony T. Yeung, Deepali Hattangadi, Lauryn Blakesley, and Emmanuelle Nicolas BioTechniques 38:749-758 (May 2005)

Mutation is as necessary for life as fidelity is in DNA replication. The study of mutations reveals the normal functions of genes, messages, proteins, the causes of many diseases, and the variability of responses among individuals. Indeed, recent mutations that have not yet become polymorphisms are often deleterious and pertinent to the disease history of afflicted individuals. This review discusses the principles behind a variety of methods for the detection of mutations and factors that should be considered in future methods design. One enzymatic approach in particular, using orthologs of the CEL I nuclease that show high specificity for all mismatches, appears to be easy and robust. Further developments of this and other methods will allow mutation detection to become an integral component of individualized medicine.

INTRODUCTION On December 12, 1946, long before the elucidation of the DNA double helix, Dr. Hermann J. Muller summarized in his Nobel lecture the contributions of the great geneticists of his time on the importance of elucidating the microscopic changes called mutations, the driving force of evolution in germ cells, the alterations that occur naturally or artificially in genes and in the newly duplicated genome (1). Sixty years later, we have a more precise description of the DNA, genes, genome, structure and function of mutations, but there is still room for improvement in the specificity, sensitivity, and cost-effectiveness of the methods of mutation detection. For example, the lack of an effective genome-wide mutation scanning procedure limits the contribution of mutation analysis even when it is clear that single nucleotide polymorphisms (SNPs) and mutations both impact the diseases of many individuals. This review discusses some of the properties of mutations relevant to the development of mutation detection technology and highlights the principles of some methods that are successful when used judiciously. We hope that it will stimulate future discovery of even better methods for mutation detection.

The most direct way to score a mutation is to read the DNA sequence, utilizing the fidelity of DNA polymerases and the Watson and Crick hydrogen bonding of