title: Resequencing array for gene variant detection in malignant hyperthermia and butyrylcholinestherase deficiency.
abstract: Malignant hyperthermia (MH) and butyrylcholinestherase (BCHE) deficiency are two relevant pharmacogenetic disorders in anesthetic practice linked with sequence variants, the former in the RyR1 and CACNA1S genes, the latter in the BCHE gene. Genotyping for known pathogenic variants in these genes is useful to help identify susceptible individuals, and others may exist but remain unknown, because full-length sequence of these genes is, in general, not investigated. To facilitate this task, we developed a resequencing DNA array, the perioperative patient safety (POPS) array, to be able to screen the entire coding sequences of the RyR1, CACNA1S and BCHE genes. MH-susceptible individuals (n = 121) identified with the in vitro contracture test, the standard diagnostic tool for MH susceptibility, were genotyped with the arrays. Compared with capillary sequencing, call rates with the arrays could achieve 100% at maximal sensitivity, although to reduce false positive rates, sensitivity was adjusted to 0.85, 0.87 and 0.66 for RyR1, CACNA1S and BCHE respectively, with overall base call specificity exceeding 99%. Detection of 29 predetermined RyR1 variants in 44 individuals was successful in 97% of the cases, among them all 16 variants of established diagnostic value. In a trial application of the arrays, 21 MH-susceptible subjects with no known RyR1 or CACNA1S variants were screened, resulting in the discovery of new variants, all confirmed by capillary sequencing. In conclusion, arrays offer an efficient high-throughput alternative for diagnostic genotyping of candidate genes affecting MH susceptibility, BCHE deficiency and other neuromuscular disorders, simultaneously enabling a comprehensive search for rare variants in these genes.
authors.affiliation: Department of Biomedicine, University Hospital Basel, Switzerland; Department Anesthesiology, University Hospital Basel, Switzerland.
authors.forename: Soledad
authors.initials: S
authors.lastname: Levano
authors.affiliation: Department of Biomedicine, University Hospital Basel, Switzerland; Department Anesthesiology, University Hospital Basel, Switzerland. Electronic address: asensio.gonzalez@unibas.ch.
authors.forename: Asensio
authors.initials: A
authors.lastname: Gonzalez
authors.affiliation: Department of Biomedicine, University Hospital Basel, Switzerland; Department Anesthesiology, University Hospital Basel, Switzerland.
authors.forename: Martine
authors.initials: M
authors.lastname: Singer
authors.affiliation: Biozentrum, Life Sciences Training Facility, University of Basel, Switzerland.
authors.forename: Philippe
authors.initials: P
authors.lastname: Demougin
authors.affiliation: University of Leipzig, Helios Kliniken Leipziger Land Leipzig, Germany.
authors.forename: Henrik
authors.initials: H
authors.lastname: Rüffert
authors.affiliation: Department of Biomedicine, University Hospital Basel, Switzerland; Department Anesthesiology, University Hospital Basel, Switzerland.
authors.forename: Albert
authors.initials: A
authors.lastname: Urwyler
authors.affiliation: Department of Biomedicine, University Hospital Basel, Switzerland; Department Anesthesiology, University Hospital Basel, Switzerland.
authors.forename: Thierry
authors.initials: T
authors.lastname: Girard
mesh_terms: D001049:Apnea / Q000235:genetics*; D002091:Butyrylcholinesterase / Q000172:deficiency* / Q000235:genetics; D015220:Calcium Channels / Q000235:genetics; D020746:Calcium Channels, L-Type; D019295:Computational Biology; D005091:Exons; D005820:Genetic Testing / Q000191:economics / Q000295:instrumentation*; D014644:Genetic Variation; D006801:Humans; D008305:Malignant Hyperthermia / Q000235:genetics*; D008661:Metabolism, Inborn Errors / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000191:economics; D019837:Ryanodine Receptor Calcium Release Channel / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: C412223:CACNA1S protein, human; D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D019837:Ryanodine Receptor Calcium Release Channel; D002091:Butyrylcholinesterase
keywords: Butyrylcholinestherase deficiency; DNA microarrays; Genetic predisposition to disease; Genetic variation; Malignant hyperthermia