Article

Original Article

Korean J Clin Pathol 2000; 20(2): 150-157

Published online April 1, 2000

Copyright © Korean Society for Laboratory Medicine.

Modified and Improved Amplification Refractory Mutation System Polymerase Chain Reaction Method for Apolipoprotein E Genotyping

김달식?이혜수?최삼임?서순팔*

Abstract

Background :The determination of apo E polymorphism through the phenotyping is not suitable for large studies in the clinical laboratories because of various problems. So apo E genotyping has been developed. We present a method of apo E genotyping using a modified amplification refractory mutation system (ARMS) technique. Method :We used four primers to detect the region containing two mutation points coding amino acid residues 112 and 158 and have developed a method of apo E genotyping by the modified ARMS technique. Apo E genotyping were performed by both the modified ARMS technique and the INNO-LiPA Apo E kit (Innogenetics, Belgium) with the following frequency distribution: ε2/2 (n=10), ε3/3 (n=15), ε4/4 (n=7), ε2/3 (n=9), ε2/4 (n=12), ε3/4 (n=13). Result :All the samples gave the correct and clear amplification patterns. Modified ARMS correctly distinguished among the six apo E genotypes. The apo E genotypes determined by both methods for every specimen studied were in complete agreement. Conclusion :This modified ARMS technique involved only two stages: PCR and agarose gel electrophoresis. Since apo E genotyping by the modified ARMS is reliable, simple to perform, less time consuming, and not expensive, we conclude that it is suitable for large sample studies in the clinical laboratories.

Keywords: Apolipoprotein E, Apo E genotyping, ARMS