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

Journal of Molecular Recognition : JMR
P NilssonP A Nygren

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.

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Citations

Jun 23, 1999·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·E C Nice, B Catimel
Dec 24, 2008·Analytical and Bioanalytical Chemistry·L G CarrascosaL M Lechuga
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Mar 3, 2005·Biosensors & Bioelectronics·Tieshan JiangMarco Mascini
Jan 25, 2005·Biophysical Journal·Danfeng YaoWolfgang Knoll
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Jan 4, 2006·Biosensors & Bioelectronics·Daniela Dell'AttiMarco Mascini
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Nov 11, 1999·Journal of Molecular Recognition : JMR·M H Van Regenmortel
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