Glioma test array for use with formalin-fixed, paraffin-embedded tissue

Array comparative genomic hybridization correlates with loss of heterozygosity and fluorescence in situ hybridization

Gayatry Mohapatra, Rebecca Betensky, Ezra R. Miller, Bjorn Carey, Leah D. Gaumont, David A. Engler, David N. Louis

Research output: Contribution to journalArticle

Abstract

Array-based comparative genomic hybridization (aCGH) is a powerful, high-throughput tool for whole genome analysis. Until recently, aCGH could only be reproducibly performed on frozen tissue samples and with significant tissue amounts. For brain tumors however, paraffin-embedded tissue blocks from small stereotactic biopsies may be the only tissue routinely available. The development of methods to analyze formalin-fixed, paraffin-embedded (FFPE) material therefore has the potential to impact molecular diagnosis in a significant way. To this end, we constructed a BAC array representing chromosomes 1, 7, 19, and X because 1p/19q deletion and EGFR gene amplification provide clinically relevant information for glioma diagnosis. We also optimized a two-step labeling procedure using an amine-modified nucleotide for generating aCGH probes. Using this approach, we analyzed a series of 28 FFPE oligodendroglial tumors for alterations of chromosomes 1, 7, and 19. We also independently assayed these tumors for 1p/19q deletion by fluorescence in situ hybridization and by loss of heterozygosity analyses. The concordance between aCGH, standard loss of heterozygosity and fluorescence in situ hybridization was nearly 100% for the chromosomes analyzed. These results suggest that aCGH could offer an improved molecular diagnostic approach for gliomas because of its ability to detect clinically relevant molecular alterations in small FFPE specimens.

Original languageEnglish (US)
Pages (from-to)268-276
Number of pages9
JournalJournal of Molecular Diagnostics
Volume8
Issue number2
DOIs
StatePublished - Jan 1 2006

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Comparative Genomic Hybridization
Loss of Heterozygosity
Fluorescence In Situ Hybridization
Glioma
Paraffin
Formaldehyde
Chromosomes, Human, Pair 7
Chromosomes, Human, Pair 1
erbB-1 Genes
Chromosomes, Human, Pair 19
Molecular Pathology
Gene Amplification
Brain Neoplasms
Amines
Neoplasms
Nucleotides
Chromosomes
Genome
Biopsy

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Medicine

Cite this

Glioma test array for use with formalin-fixed, paraffin-embedded tissue : Array comparative genomic hybridization correlates with loss of heterozygosity and fluorescence in situ hybridization. / Mohapatra, Gayatry; Betensky, Rebecca; Miller, Ezra R.; Carey, Bjorn; Gaumont, Leah D.; Engler, David A.; Louis, David N.

In: Journal of Molecular Diagnostics, Vol. 8, No. 2, 01.01.2006, p. 268-276.

Research output: Contribution to journalArticle

Mohapatra, Gayatry ; Betensky, Rebecca ; Miller, Ezra R. ; Carey, Bjorn ; Gaumont, Leah D. ; Engler, David A. ; Louis, David N. / Glioma test array for use with formalin-fixed, paraffin-embedded tissue : Array comparative genomic hybridization correlates with loss of heterozygosity and fluorescence in situ hybridization. In: Journal of Molecular Diagnostics. 2006 ; Vol. 8, No. 2. pp. 268-276.
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