Identification of specific BRCA1 and BRCA2 variants by DHPLC

Hum Mutat. 2000 Oct;16(4):345-53. doi: 10.1002/1098-1004(200010)16:4<345::AID-HUMU7>3.0.CO;2-#.


Denaturing high performance liquid chromatography (DHPLC) is generating increasing interest in clinical genetics as a reliable tool for the analysis of genetic alterations. In the work presented here our intentions were to optimize primer design and DHPLC analysis conditions for a qualitative detection of BRCA1 and BRCA2 variations. The BRCA1 and BRAC2 genes display a high proportion of polymorphisms. Sequencing efforts geared towards the distinction of tumor-related mutations and benign variants still remain time-consuming and expensive. DHPLC elution profiles, however, permit the correlation of a characteristic chromatographic profile with a specific sequence alteration. In this study we evaluate the sensitivity of DHPLC for the identification of unique polymorphisms, which are frequent in the Caucasian population, in lieu of sequence analysis. The complete BRCA1 gene and parts of BRCA2 were examined. In the case of BRCA1, 431 out of 432 heterozygotes were identified correctly. In addition, 18 new profiles were identified which had not been detected previously in our studies and which represented new mutations or rare polymorphisms. For BRCA2, 135 out of 137 simple sequence variants were classified correctly. In addition, six new profiles were identified which represented new mutations or rare polymorphisms.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • BRCA2 Protein
  • Base Sequence
  • Breast Neoplasms / chemistry
  • Breast Neoplasms / genetics
  • Chromatography, High Pressure Liquid / methods*
  • Genes, BRCA1 / genetics*
  • Genetic Markers / genetics
  • Humans
  • Molecular Sequence Data
  • Neoplasm Proteins / genetics*
  • Neoplasm Proteins / isolation & purification
  • Nucleic Acid Denaturation / genetics*
  • Polymorphism, Genetic
  • Protein Isoforms / genetics
  • Protein Isoforms / isolation & purification
  • Transcription Factors / genetics*
  • Transcription Factors / isolation & purification


  • BRCA2 Protein
  • Genetic Markers
  • Neoplasm Proteins
  • Protein Isoforms
  • Transcription Factors