Evaluation of arrayed primer extension for TP53 mutation detection in breast and ovarian carcinomas
| Autor: | Eldri U. Due, Elda Tagliabue, Jahn M. Nesland, Anne Lise Børresen-Dale, Pedro Kringen, Aune Nehman, Neeme Tõnisson, Yun Wang, Anna Bergamaschi |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Electrophoresis
Time Factors Microarray DNA Mutational Analysis Breast Neoplasms DNA Fragmentation Computational biology Biology medicine.disease_cause Polymerase Chain Reaction General Biochemistry Genetics and Molecular Biology Primer extension Exon Cell Line Tumor Genotype medicine Humans Missense mutation Codon Genotyping Alleles DNA Primers Oligonucleotide Array Sequence Analysis Ovarian Neoplasms Genetics Mutation Sequence Analysis DNA Oligonucleotides Antisense Genes p53 Immunohistochemistry Female Tumor Suppressor Protein p53 Primer (molecular biology) Biotechnology |
| Zdroj: | BioTechniques. 39:755-761 |
| ISSN: | 1940-9818 0736-6205 |
| Popis: | Mutations in the tumor suppressor gene TP53 are associated with a wide range of different cancers and may have prognostic and therapeutic implications. Methods for rapid and sensitive detection of mutations in this gene are therefore required. In order to make screening more effective, a commercially available TP53 genotyping microarray from Asper Biotech has been constructed by arrayed primer extension (APEX). The present study is the first report that blindly evaluates the efficiency of the second generation APEX TP53 genotype chip outside the Asper laboratory and compares it to temporal temperature gradient electrophoresis (TTGE) and sequencing of TP53 for mutation detection in ovarian and breast cancer samples. All nucleotides in the TP53 gene from exon 2–9 are included on the chip by synthesis and application of sequence-specific oligonucleotides. The chip was validated by screening 48 breast and 11 ovarian cancer cases, all of which had previously been analyzed by TTGE and sequencing. APEX scored 17 of 20 sequence variants, missing one deletion, one insertion, and a missense mutation. Resequencing efficiency using APEX was 92% for both DNA strands and 99.5% for sense and/or antisense strand. We conclude that the APEX TP53 microarray is a robust, rapid, and comprehensive screening tool for sequence alterations in tumors. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|