Automated array-CGH optimized for archival formalin-fixed, paraffin-embedded tumor material

BMC Cancer. 2007 Mar 7;7:43. doi: 10.1186/1471-2407-7-43.

Abstract

Background: Array Comparative Genomic Hybridization (aCGH) is a rapidly evolving technology that still lacks complete standardization. Yet, it is of great importance to obtain robust and reproducible data to enable meaningful multiple hybridization comparisons. Special difficulties arise when aCGH is performed on archival formalin-fixed, paraffin-embedded (FFPE) tissue due to its variable DNA quality. Recently, we have developed an effective DNA quality test that predicts suitability of archival samples for BAC aCGH.

Methods: In this report, we first used DNA from a cancer cell-line (SKBR3) to optimize the aCGH protocol for automated hybridization, and subsequently optimized and validated the procedure for FFPE breast cancer samples. We aimed for highest throughput, accuracy, and reproducibility applicable to FFPE samples, which can also be important in future diagnostic use.

Results: Our protocol of automated array-CGH on archival FFPE ULS-labeled DNA showed very similar results compared with published data and our previous manual hybridization method.

Conclusion: This report combines automated aCGH on unamplified archival FFPE DNA using non-enzymatic ULS labeling, and describes an optimized protocol for this combination resulting in improved quality and reproducibility.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Automation
  • Biopsy, Needle
  • Breast Neoplasms / genetics*
  • Breast Neoplasms / pathology
  • Cell Line, Tumor
  • DNA, Neoplasm / analysis*
  • Female
  • Formaldehyde / pharmacology
  • Humans
  • Nucleic Acid Hybridization / methods*
  • Oligonucleotide Array Sequence Analysis / methods*
  • Paraffin Embedding
  • Reproducibility of Results
  • Sampling Studies
  • Sensitivity and Specificity
  • Tissue Fixation / methods

Substances

  • DNA, Neoplasm
  • Formaldehyde