title: Pitfalls in LightCycler diagnosis of the single-nucleotide polymorphism 13.9 kb upstream of the lactase gene that is associated with adult-type hypolactasia.
abstract: BACKGROUND
  Patients presenting with symptoms of lactose intolerance are in some centres routinely tested for a single-nucleotide polymorphism C-13910T, which is located upstream of the lactase gene (LCT) and is tightly associated with genetically determined lactase persistence/non-persistence. Typing of this polymorphism enables differential diagnosis for genetic versus secondary causes of lactose intolerance. Several PCR-based methods have been established as tests for this SNP. In particular, automated genotyping assays conducted on LightCycler platforms provide a rapid, labour-saving means for routine high-throughput analysis of this variant. Recently, several novel allelic variants have been identified in non-European populations. Three of these variants occur in close proximity to C-13910T, but their effect on the genetic test is unknown.
  
  
  METHODS
  Here we analyse whether the occurrence of C-13907G, T-13913C, and T-13915G, affect the diagnostic accuracy of C-13910T typings obtained using the LightCycler MutaREAL lactase real-time PCR kit.
  
  
  RESULTS
  Genotyping of DNA samples harbouring respective variants or combinations thereof significantly influenced the LightCycler analysis. Some allelic combinations generated melting profiles that prevented the correct assignment of C-13910T.
  
  
  CONCLUSIONS
  We conclude that genotyping of the C-13910T variant with the MutaREAL lactase real-time PCR kit in non-Europeans is prone to error and should be omitted.
authors.affiliation: Institute of Clinical Chemistry and Pathobiochemistry, RWTH-University Hospital, D-52074 Aachen, Germany. rweiskirchen@ukaachen.de
authors.forename: Ralf
authors.initials: R
authors.lastname: Weiskirchen
authors.forename: Carmen G
authors.initials: CG
authors.lastname: Tag
authors.forename: Senait
authors.initials: S
authors.lastname: Mengsteab
authors.forename: Axel M
authors.initials: AM
authors.lastname: Gressner
authors.forename: Catherine J E
authors.initials: CJ
authors.lastname: Ingram
authors.forename: Dallas M
authors.initials: DM
authors.lastname: Swallow
mesh_terms: D000328:Adult; D005838:Genotype; D006801:Humans; D043322:Lactase / Q000235:genetics*; D007787:Lactose Intolerance / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA
chemical_list: D011933:Reagent Kits, Diagnostic; D043322:Lactase