Rapid Apolipoprotein E Genotyping by the Multiplex Amplification Refractory Mutation System

Lee, Mi Kyung; Park, Ae Ja

Abstract

BACKGROUND
Currently, several different apolipoprotein E (apo E) genotyping methods have been developed. The Amplification Refractory Mutation System (ARMS) apo E genotyping, as previously described, requires four separate PCR reactions. The purpose of this study is to determine the clinical usefulness of the multiplex ARMS apo E genotyping with the use of only two PCR reactions.
METHODS
We used five primers and two separate PCR reactions to detect the apo E polymorphism by using the multiplex ARMS technique. Apo E genotyping was performed with both the multiplex ARMS and INNO-LiPATM Apo E kit (INNOGENETICS) in 122 random samples. We investigated the effect of dimethyl sulfoxide (DMSO) in the multiplex ARMS PCR with various DMSO concentrations (0-15%).
RESULTS
All six possible genotypes for apo E were clearly discernible with the multiplex ARMS. The apo E genotypes determined by the two methods were in complete agreement with all 122 samples. We found that DMSO is essential for the successful amplification of the multiplex ARMS and DMSO at concentrations of 3%-7% to be the optimal concentration.
CONCLUSIONS
ARMS analysis involves two stages: PCR and agarose gel electrophoresis. Apo E genotyping using the multiplex ARMS requires only two PCR reactions. Thus, because of its simplicity, speed, accuracy, and cost-effectiveness, this method may be appropriate for determining the apo E genotypes in routine clinical laboratories.