Identification of apolipoprotein E polymorphism by using synthetic oligonucleotides.

Abstract

A procedure based on selective hybridization with allele-specific oligonucleotides was developed for typing apolipoprotein E variants from human genomic DNAs. Two sets of oligonucleotides were synthesized and used to discriminate either between epsilon 3 and epsilon 4 alleles or between epsilon 3 and epsilon 2 alleles. Combination of the allele-specific oligonucleotide hybridization with the method for in vitro DNA amplification (Polymerase Chain Reaction) Saiki, R. K. et al. 1985. Science, 230: 1350-1354) (1) dramatically improved the sensitivity and the reliability of the procedure. Adaptation of a simple strategy involving direct cloning and DNA sequencing of in vitro amplified DNA enables rapid identification of any mutation within the apoE gene area encoding the receptor binding domain.