title: Simple and precise detection of UGT1A1 polymorphisms with a modified loop-hybrid mobility shift assay using Cy5-labeled loop probes.
abstract: BACKGROUND
  Irinotecan, an inhibitor of topoisomerase I, has been widely used as an important anti-cancer therapeutic drug. Deleterious effects of the drug in hypersensitive patients are known to be associated with genetic polymorphisms of the UGT1A1 gene, namely the polymorphic variants, *28 and *6.
  
  
  METHODS
  A modified form of loop-hybrid mobility shift assay using a Cy5-tagged loop-hybrid probe was proposed as a precise and easy method of determining TA repeat polymorphisms at the *28 locus.
  
  
  RESULTS
  In this modified method, only loop-hybrid bands were detected by a Cy5-fluorescent signal, despite several irregular electrophoretic bands due to TA repeats in the PCR product.
  
  
  CONCLUSIONS
  When a loop-hybrid using a Cy5-tagged probe for the *28 locus and *6 locus were combined and used for mobility shift assay, simultaneous typing of the *28 and *6 variants was achieved in a single lane.
authors.affiliation: Molecular Pathology and Genetics Division, Kanagawa Cancer Center Research Institute, Asahi-ku, Yokohama, Japan.
authors.forename: Shoichi
authors.initials: S
authors.lastname: Matsukuma
authors.forename: Haruhiro
authors.initials: H
authors.lastname: Saito
authors.forename: Kozo
authors.initials: K
authors.lastname: Yamada
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Okamoto
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Ishikawa
authors.forename: Mitsuyo
authors.initials: M
authors.lastname: Yoshihara
authors.forename: Shiro
authors.initials: S
authors.lastname: Koizume
authors.forename: Yuji
authors.initials: Y
authors.lastname: Sakuma
authors.forename: Yohei
authors.initials: Y
authors.lastname: Miyagi
mesh_terms: D005838:Genotype; D014453:Glucuronosyltransferase / Q000235:genetics*; D006801:Humans; D015335:Molecular Probes; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D000099148:UGT1A1 Enzyme
chemical_list: D014453:Glucuronosyltransferase; D015335:Molecular Probes; D000099148:UGT1A1 Enzyme