title: Rapid simultaneous genotyping of the frequent butyrylcholinesterase variants Asp70Gly and Ala539Thr with fluorescent hybridization probes.
abstract: BACKGROUND
  The clinically important variants of butyrylcholinesterase (BChE) are the A- (Asp70Gly) and K-variants (Ala539Thr), which are common among Caucasians. These variants are associated with abnormal drug metabolism during anaesthesia, which leads to a prolonged neuromuscular block following administration of the neuromuscular blocking agents, mivacurium and succinylcholine. In addition, the K-variant has been proposed to be associated with Alzheimer's disease together with apolipoprotein E epsilon4. To facilitate diagnostics, we set out to establish a rapid and simple method for simultaneous genotyping of the A- and K-variants.
  
  
  METHODS
  Using the LightCycler, a rapid-cycle duplex PCR is combined with generation of allele-specific fluorescent probe melting profiles. This allows simultaneous detection of both of the mutations in the BChE gene. The results were compared with direct sequencing and phenotyping results.
  
  
  RESULTS
  Samples from 80 subjects were genotyped. The genotypes determined using the LightCycler were identical to those obtained by direct sequencing of conventional polymerase chain reaction products and was more accurate than phenotyping based on biochemical assays.
  
  
  CONCLUSIONS
  A high-speed and easy to perform mutation detection assay has been established for the two most common mutations, Asp70Gly and Ala539Thr, in BChE, using the LightCycler technology and melting curves.
authors.affiliation: Department of Anaesthesia, Center of Head and Orthopaedics, Rigshospitalet, Denmark. mgatke@rh.dk
authors.forename: M R
authors.initials: MR
authors.lastname: Gätke
authors.forename: J
authors.initials: J
authors.lastname: Viby-Mogensen
authors.forename: J R
authors.initials: JR
authors.lastname: Bundgaard
mesh_terms: D000328:Adult; D019943:Amino Acid Substitution / Q000235:genetics*; D001483:Base Sequence; D002091:Butyrylcholinesterase / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods / Q000592:standards; D005453:Fluorescence; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010641:Phenotype; D015203:Reproducibility of Results
chemical_list: D002091:Butyrylcholinesterase