Rapid and accurate detection of atypical and Kalow variants in the butyrylcholinesterase gene using denaturing high performance liquid chromatography.
BACKGROUND
Variations in the butyrylcholinesterase (BCHE) gene can prolong neuromuscular block after the administration of the neuromuscular blocking drugs succinylcholine and mivacurium. The most frequent variations in the BCHE gene are the atypical (A) and Kalow (K) variants. We, therefore, developed a detection approach for these most common variants based on the method of denaturing high-performance liquid chromatography (dHPLC).


METHODS
Forty-six subjects were included. Sixteen were known to carry either the normal sequence or different combinations of A- and K-variants and 30 test subjects were randomly selected from the general population. Three samples of the 16 were used to establish the dHPLC protocols. Results from dHPLC were blindly compared with the automated sequencing data.


RESULTS
All A- and K-variants in their heterozygous and homozygous forms were unequivocally identified by dHPLC. For all 86 tested alleles, dHPLC results were 100% concordant with DNA sequencing results. Additional genetic variations were also detected, which included G344A encoding for the silent variant (S) and a known polymorphism, A1914G.


CONCLUSIONS
The described dHPLC protocols offer a rapid and accurate screening method for the most common variants of BCHE. This screening tool might be useful in pharmacokinetic studies investigating drugs metabolized by BCHE, such as mivacurium and succinylcholine.
Levano
Soledad
S
Department of Anesthesia and Research, University of Basel, Basel, Switzerland. s.levano@unibas.ch
Keller
Dagmar
D
Schobinger
Elektra
E
Urwyler
Albert
A
Girard
Thierry
T
D002091:Butyrylcholinesterase / Q000097:blood / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004252:DNA Mutational Analysis; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D009691:Nucleic Acid Denaturation; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results
D002091:Butyrylcholinesterase