Increased Sensitivity for Detection of Mutations in Exon 15 of the APC
Gene in Patients with Familial Adenomatous Polyposis
Konvalinka D., 1Bik E. C., 1Tops C. M., 1Bakker E., Singh A. R., Vrtěl R., Šantavý J.
Ústav lékařské genetiky a fetální medicíny LF UP a FN, Olomouc 1Center for Human and Clinical Genetics, LUMC, Leiden, The Netherlands
Background. Protein Truncation Test (PTT) was used to detect mutations in exon 15 of the APC gene in patients
with Familial Adenomatous Polyposis. This method is limited by its ability to detect polypetide chains up to a certain
minimum length. The aim of this study was to increase the sensitivity of detection of mutations in this region by
using the technique of Denaturing Gradient Gel Electrophoresis (DGGE).
Methods and Results. Study were performed on 122 patients without detected mutations in the APC gene. The
patients were divided into two independent groups 15A and 15A+15B (with 51 and 71 patients respectively). All
the patients were tested with the DGGE and the positive findings were confirmed with sequencing. No mutation was
detected in the group 15A (0 %). In group 15A+15B one (1,4 %) polymorphism and four (5,63 %) patients with
nonsense mutations were detected.
Conclusions. DGGE is an effective method for detecting mutations in the first part of exon 15 of APC gene. It allows
detecting any change in DNA strand. DGGE complements PTT in scanning of the whole exon 15 of APC gene.
familial adenomatous polyposis, adenomatous polyposis coli, APC gene, denaturing gradient gel