title: Length analysis of polymerase chain reaction products: a sensitive and reliable technique for the detection of mutations in KIT exon 11 in gastrointestinal stromal tumors.
abstract: Gastrointestinal stromal tumors are the most common mesenchymal neoplasms of the digestive tract. These tumors express the c-kit receptor tyrosine kinase, and many have activating mutations in the juxtamembrane region coded by the exon 11 of KIT. Detection of these mutations has prognostic and therapeutic impact. The aim of the study was to compare a new detection method by length analysis of polymerase chain reaction products (LAPP) to direct sequencing. The detection of either deletion or insertion mutations within the exon 11 of KIT was performed on genomic DNA extracted from 40 paraffin-embedded samples from 38 patients. Double-strand direct sequencing revealed a mutation in 25 of 40 samples. In two additional samples, a mutation was suspected but could not be determined by sequencing. LAPP revealed a mutation in 27 samples, corresponding to the 25 determined and 2 suspected samples. One of these latter samples contained three different alleles. Mutations corresponded to either deletions (n = 24) or insertion (n = 1) and had the same size with sequencing and LAPP. Our results show that LAPP is as accurate and more sensitive than direct sequencing for the detection of deletion or insertion mutations of exon 11 of KIT in gastrointestinal stromal tumors.
authors.affiliation: Service d'anatomie Pathologique, Hôpital Paul Brousse and UPRES 1596, Université Paris Sud, France. jean-francois.emile@pbr.ap-hop-paris.fr
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Emile
authors.forename: Antoinette
authors.initials: A
authors.lastname: Lemoine
authors.forename: Nathalie
authors.initials: N
authors.lastname: Bienfait
authors.forename: Philippe
authors.initials: P
authors.lastname: Terrier
authors.forename: Daniel
authors.initials: D
authors.lastname: Azoulay
authors.forename: Brigitte
authors.initials: B
authors.lastname: Debuire
mesh_terms: D001483:Base Sequence; D004252:DNA Mutational Analysis; D005091:Exons / Q000235:genetics*; D005770:Gastrointestinal Neoplasms / Q000737:chemistry / Q000235:genetics* / Q000473:pathology; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D019009:Proto-Oncogene Proteins c-kit / Q000032:analysis / Q000235:genetics*; D015203:Reproducibility of Results; D012509:Sarcoma / Q000737:chemistry / Q000235:genetics* / Q000556:secondary; D012680:Sensitivity and Specificity; D017154:Stromal Cells / Q000737:chemistry / Q000473:pathology
chemical_list: D019009:Proto-Oncogene Proteins c-kit