Detection of mutations in PCR products from clinical samples by surface plasmon resonance

✍ Scribed by Peter Nilsson; Björn Persson; Anita Larsson; Mathias Uhlén; Per-Åke Nygren

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 330 KB
• Volume: 10
• Category: Article
• ISSN: 0952-3499
• DOI: 10.1002/(sici)1099-1352(199701/02)10:1<7::aid-jmr341>3.0.co;2-9

✦ Synopsis

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.