Volume 3 Supplement 1

23rd Congress of the International Association for Breast Cancer Research

Open Access

Mutation detection in familial and sporadic breast cancers by denaturing high-performance liquid chromatography (DHPLC)

  • B Betz1,
  • D Larbig1,
  • TO Goecke1,
  • C Nestle-Krämling1,
  • HG Bender1 and
  • D Niederacher1
Breast Cancer Research20013(Suppl 1):A9

https://doi.org/10.1186/bcr409

Received: 10 May 2001

Published: 31 May 2001

Objective

Denaturing high-performance liquid chromatography (DHPLC) is a recently developed method for detection of mutation that is gaining importance as a screening method for analyzing familial breast cancers, as well as heterogeneous tumor material.

Method

DHPLC was established for mutation detection in BRCA1/2 diagnostic, using more than 200 different positive controls. Up until now, 64 DNA samples from patients with familial background for breast cancer (BC) were analyzed by DHPLC for BRCA1/2 mutations. An additional 136 sporadic BC were examined for p53 mutations, analyzing exons 5-8 by DHPLC. Positive results were confirmed by direct DNA sequencing.

Results

The analysis of 64 DNA samples from patients with familial background for BC revealed several mutations and unclassified variants (UVs). Twenty-three different p53 mutations could be detected in 138 sporadic BC. Dilution of mutant DNA by wild-type DNA revealed the high sensitivity of this method: 5% mutant DNA is sufficient to achieve a positive DHPLC result. However, confirming a positive DHPLC result by DNA sequencing is difficult in heterogeneous tumor material.

Conclusion

DHPLC is a reliable, high-throughput technique for detection of mutation in familial breast cancers, as well as in heterogeneous tumor material.

Authors’ Affiliations

(1)
Department of Obstetrics & Gynecology, Heinrich-Heine-University

Copyright

© BioMed Central Ltd 2001

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