NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis

High-resolution copy number analysis of paraffin-embedded archival tissue using SNP BeadArrays.

Author(s): Oosting J, Lips EH, van Eijk R, Eilers PH, Szuhai K, Wijmenga C, Morreau H, van Wezel T

Publication: Genome Res, 2007, Vol. 17, Page 368-76

PubMed ID: 17267813 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of using formalin-fixed paraffin-embedded (FFPE) instead of frozen specimens on copy number determination by array comparative genomic hybridization (aCGH) and to compare the results between three array types.

Conclusion of Paper

All three platforms slightly overestimated the copy number for normal specimens, and copy numbers for regions with a gain were slightly higher while those with a loss were slightly lower when determined by GeneChip than when determined with bacterial artificial chromosome (BAC) arrays or BeadArrays. The copy number variation (CNV) was comparable between frozen and FFPE specimens using BAC and BeadArrays. The authors report the BeadArrays allowed for detection of loss of heterozygosity (LOH) by physical loss as well as copy neutral LOH.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effects of using FFPE instead of frozen colorectal tumors and normal adjacent specimens on copy number determination by aCGH and to compare the results obtained between three array types. DNA was extracted from sections of frozen specimens using the genomic Wizard kit and from 0.6 mm punches of FFPE tissue using Chelex-100 and cleaned up with the Wizard kit.

    Summary of Findings:

    All three platforms slightly overestimated the copy number for normal specimens, and copy numbers for regions with a gain were slightly higher while those with a loss were slightly lower when determined by GeneChip rather than with BAC arrays or BeadArrays. The CNV was comparable between frozen and FFPE specimens using BAC and BeadArrays, but the correlation between frozen and FFPE specimens was 0.85 which was lower than the correlations between arrays (r=0.93-0.95) or between replicates with different intensity (r=0.92). However, exclusion of one specimen increased the correlation between FFPE and frozen specimens using BeadArrays to 0.96. The authors report the BeadArrays allowed for detection of LOH by physical loss as well as copy neutral LOH.

    Biospecimens
    Preservative Types
    • Formalin
    • Frozen
    Diagnoses:
    • Neoplastic - Carcinoma
    • Neoplastic - Normal Adjacent
    • Neoplastic - Benign
    Platform:
    AnalyteTechnology Platform
    DNA Array CGH
    DNA DNA microarray
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Type of fixation/preservation Formalin (buffered)
    Frozen
    Array CGH Specific Type of array Illumina Sentrix Beadarray
    Affymetrix 10 K Xba1 2.0 array
    Custom BAC array

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