title: Single-nucleotide polymorphisms may cause erroneous results in primer-introduced restriction enzyme analyses: a case of molecular misdiagnosis of homozygous vs heterozygous familial hypercholesterolemia.
abstract: PCR amplification followed by a primer introduced restriction analysis PCR (PIRA-PCR) is a widely used method to detect point mutations. Usually the artificial RFLP is created by siting one nucleotide mismatch near the 3; end of the primer. This does not alter the hybrization of the primer to the target DNA sequence. Unfortunately, unexpected single nucleotide polymorphisms (SNPs) may lead to additional mismatches and result in no amplification of the allele having unexpected SNP. We describe a warning example in which heterozygous familial hypercholesterolemia patient had an unexpected SNP and this led to his misdiagnosis.
authors.affiliation: Department of Medicine, University of Helsinki, Helsinki, FIN-00290, Finland. Alpo.Vuorio@huch.fi
authors.forename: A F
authors.initials: AF
authors.lastname: Vuorio
authors.forename: L
authors.initials: L
authors.lastname: Paulin
authors.forename: J
authors.initials: J
authors.lastname: Saltevo
authors.forename: K
authors.initials: K
authors.lastname: Kontula
mesh_terms: D020137:Base Pair Mismatch; D017931:DNA Primers; D003951:Diagnostic Errors / Q000379:methods; D006580:Genetic Carrier Screening / Q000379:methods*; D006720:Homozygote; D006801:Humans; D006938:Hyperlipoproteinemia Type II / Q000175:diagnosis* / Q000235:genetics*; D019656:Loss of Heterozygosity / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D015203:Reproducibility of Results; D015183:Restriction Mapping / Q000379:methods*
chemical_list: D017931:DNA Primers