title: Gene analysis of genomic DNA from stored serum by polymerase chain reaction: identification of three missense mutations in patients with cholinesterasemia and ABO genotyping.
abstract: We established a method to determine the butyrylcholinesterase genotype associated with a BCHE deficiency directly using multiple PCR from stored serum, which was stored at -70 degrees C for more than 30 years. PCR products from sera of six propositi were used for DNA sequence analysis. All of these BChE variants were characterized by a single nucleotide substitution. Four of them were homozygotes and demonstrated a C-->T single nucleotide point mutation at codon 100 from CCA (Pro) to TCA (Ser). The fifth case was a heterozygote of this mutation. The remaining one was a compound heterozygote showing a T-->C transition mutation at codon 203 from TCA (Ser) to CCA (Pro) and a G-->C transversion mutation at codon 365 from GGA (Gly) to CGA (Arg). Furthermore we developed a method to determine the ABO genotype from the same serum. These results indicated that serum is useful as a starting material for amplification of genomic DNA when fresh blood samples are not available.
authors.affiliation: Department of Biochemistry, Kawasaki Medical School, 577 Matsushima, Kurashiki 701-0192, Japan. kazuohi@med.kawasaki-m.ac.jp
authors.forename: K
authors.initials: K
authors.lastname: Hidaka
authors.forename: Y
authors.initials: Y
authors.lastname: Watanabe
authors.forename: M
authors.initials: M
authors.lastname: Tomita
authors.forename: N
authors.initials: N
authors.lastname: Ueda
authors.forename: M
authors.initials: M
authors.lastname: Higashi
authors.forename: Y
authors.initials: Y
authors.lastname: Minatogawa
authors.forename: I
authors.initials: I
authors.lastname: Iuchi
mesh_terms: D000017:ABO Blood-Group System / Q000235:genetics*; D001483:Base Sequence; D002091:Butyrylcholinesterase / Q000097:blood*; D004247:DNA / Q000097:blood / Q000235:genetics*; D017931:DNA Primers; D005260:Female; D005838:Genotype; D006801:Humans; D008297:Male; D020125:Mutation, Missense; D010375:Pedigree; D016133:Polymerase Chain Reaction
chemical_list: D000017:ABO Blood-Group System; D017931:DNA Primers; D004247:DNA; D002091:Butyrylcholinesterase