Caister Academic Press

The Quantitative Amplification Refractory Mutation System

Authors: P. Punia and N. Saunders
Abstract: The amplification refractory mutation system (ARMS), which has also been described as allele-specific PCR (ASP) and PCR amplification of specific alleles (PASA), is a PCR-based method of detecting single base mutations. ARMS has been applied successfully to the analysis of a wide range of polymorphisms, germ- line mutations and somatic mutations. The technique has the ability to discriminate low-levels of the mutant sequence in a high background of wild-type DNA. In an ARMS PCR the terminal 3'nucleotide of one of the PCR primers coincides with the target mutation. Most applications of the method rely on end-point analysis, utilising the classic gel-electrophoresis method. However, end-point analysis can only assess the presence or absence of mutant or wild-type sequences and does not give an indication of the ratio of mutant to wild-type in a mixed population of DNA. Here we describe a real-time PCR adaptation of ARMS, quantitative ARMS, that allows measurement of the size of the population of each variant in a mixture. A method for the detection of human hepatitis B virus mutations that confer resistance to the antiviral lamivudine is described as an example.
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