Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS)

Abstract

We have improved the "polymerase chain reaction" (PCR) to permit rapid analysis of any known mutation in genomic DNA. We demonstrate a system, ARMS (Amplification Refractory Mutation System), that allows genotyping solely by inspection of reaction mixtures after agarose gel electrophoresis. The system is simple, reliable and non-isotopic. It will clearly distinguish heterozygotes at a locus from homozygotes for either allele. The system requires neither restriction enzyme digestion, allele-specific oligonucleotides as conventionally applied, nor the sequence analysis of PCR products. The basis of the invention is that unexpectedly, oligonucleotides with a mismatched 3'-residue will not function as primers in the PCR under appropriate conditions. We have analysed DNA from patients with alpha 1-antitrypsin (AAT) deficiency, from carriers of the disease and from normal individuals. Our findings are in complete agreement with allele assignments derived by direct sequencing of PCR products.

Overview of molecular testing in non-small-cell lung cancer: mutational analysis, gene copy number, protein expression and other biomarkers of EGFR for the prediction of response to tyrosine kinase inhibitors

“…These methods allow the rapid mutation screening of large numbers of samples with increased sensitivity but still require direct sequencing to identify or confirm mutations. The amplification refractory mutation system (ARMS) (Newton et al, 1989;Whitcombe et al, 1999), combined with the scorpion method (Kimura et al, 2006) and the peptide nucleic acid-locked clamping method (Nagai et al, 2005), are alternative sensitive methods that may be used to detect rapidly specific EGFR mutations using real-time PCR-based technology.…”

Prenatal Diagnosis of Sickle Cell Disease by the Technique of PCR

“…Then a modification of PCR technique, known as Amplification Refractory Mutation System (ARMS), was developed by Newton et al [12] in 1989 and subsequently ARMS-PCR was used for the prenatal diagnosis of SCD [13][14][15], since it is a simple technique in comparison to other PCR based techniques and provide rapid results even for large number of samples [16].…”

Molecular genetic confirmatory testing from newborn screening samples for the common African‐American, Asian Indian, Southeast Asian, and Chinese β‐thalassemia mutations

“…Most of the molecular diagnostic laboratories utilize PCR amplification, with allele-specific amplification using an amplification refractory mutation system (ARMS), allele-specific oligonucleotide probes (ASO), or reverse dot blot methodology to define the known mutations that cause b-thalassemia [11][12][13]. However, for unknown mutations of the b-globin gene, single-stranded confirmation polymorphism (SSCP) and denaturing gradient gel electrophoresis (DGGE) may be used [13][14][15].…”