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

J Mol Recognit. 1997 Jan-Feb;10(1):7-17. doi: 10.1002/(SICI)1099-1352(199701/02)10:1<7::AID-JMR341>3.0.CO;2-9.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry
  • Biosensing Techniques
  • Biotin / chemistry
  • Breast Neoplasms / genetics
  • Chemistry, Clinical / instrumentation
  • Chemistry, Clinical / methods*
  • DNA Mutational Analysis / methods*
  • Genes, p53
  • Humans
  • In Situ Hybridization / methods
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Streptavidin
  • Surface Properties
  • Tumor Suppressor Protein p53 / genetics*

Substances

  • Bacterial Proteins
  • Tumor Suppressor Protein p53
  • Biotin
  • Streptavidin