title: Detection of mutations in PCR products from clinical samples by surface plasmon resonance.
abstract: Two different strategies for scanning and screening of mutations in polymerase chain reaction (PCR) products by hybridization analysis are described, employing real-time biospecific interaction analysis (BIA) for detection. Real-time BIA was used to detect differences in hybridization responses between PCR products and different 17-mer oligonucleotide probes. For the analysis using a biosensor instrument, two different experimental formats were investigated based on immobilization of either biotinylated PCR products or oligonucleotide probes onto a sensor chip. Applied on the human tumour suppressor p53 gene, differences in hybridization levels for full-match and mismatch situations employing both formats allowed the detection of point mutations in exon 6 PCR products, derived from a breast tumour biopsy sample. In addition, a mutant sample sequence could be detected in a 50/50 background of wild type exon 6 sequence. The suitability of the different formats for obtaining a regenerable system and a high throughput of samples is discussed.
authors.affiliation: Department of Biochemistry and Biotechnology, KTH-Royal Institute of Technology, Stockholm, Sweden.
authors.forename: P
authors.initials: P
authors.lastname: Nilsson
authors.forename: B
authors.initials: B
authors.lastname: Persson
authors.forename: A
authors.initials: A
authors.lastname: Larsson
authors.forename: M
authors.initials: M
authors.lastname: Uhlén
authors.forename: P A
authors.initials: PA
authors.lastname: Nygren
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D015374:Biosensing Techniques; D001710:Biotin / Q000737:chemistry; D001943:Breast Neoplasms / Q000235:genetics; D002624:Chemistry, Clinical / Q000295:instrumentation / Q000379:methods*; D004252:DNA Mutational Analysis / Q000379:methods*; D016158:Genes, p53; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019809:Streptavidin; D013499:Surface Properties; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D016159:Tumor Suppressor Protein p53; D001710:Biotin; D019809:Streptavidin