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
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Quality control and sensitivity of polymerase chain reaction techniques for the assessment of immunoglobulin heavy chain gene rearrangements from fixed- and paraffin-embedded samples.

Author(s): Camilleri-Broët S, Devez F, Tissier F, Ducruit V, Le Tourneau A, Diebold J, Audouin J, Molina T

Publication: Ann Diagn Pathol, 2000, Vol. 4, Page 71-6

PubMed ID: 10760319 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of fixative type and fixation duration on detection of immunoglobulin H (IgH) rearrangements in lymphomas by PCR.

Conclusion of Paper

Rearrangement of the framework region (FR)3 locus was found in 14 of 20 frozen lymphoma specimens. The rearrangements were confirmed for 13 of the 14 cases when specimens were fixed in formalin or Bouin's fixative. In the 14'th specimen, only the B-globin control was successfully amplified. In the single case of splenic mantle cell lymphoma, all three FR rearrangements were found and could be detected equally well in specimens that were frozen, fixed in formalin for up to 15 h, or fixed in Bouin's fixative for up to 4 h. However, only approximately 10% of FR3 and B-globin and none of the other markers were detectable when formalin-fixed specimens were incubated for 48 h, as opposed to overnight, in a vacuum infiltration processor or when fixed in Bouin's fixative for 6 h.

Studies

  1. Study Purpose

    The purpose of this study was to compare detection of IgH rearrangements by PCR in frozen lymphomas versus lymphomas fixed in formalin for 8 h-15 days or fixed in Bouin's fixative for 1-50 h.

    Summary of Findings:

    Rearrangement of the FR3 locus was found in 14 of 20 frozen lymphoma specimens. The rearrangements were confirmed for 13 of the 14 cases when specimens were fixed in formalin or Bouin's fixative. In the 14'th specimen, only the B-globin control was successfully amplified. In the single case of splenic mantle cell lymphoma, all three FR rearrangements were found and could be detected equally well in specimens that were frozen, fixed in formalin for up to 15 days, or fixed in Bouin's fixative for up to 4 h. However, only approximately 10% of FR3 and B-globin and none of the other markers were detectable when formalin-fixed specimens were incubated for 48 h, as opposed to overnight, in a vacuum infiltration processor or when fixed in Bouin's fixative for 6 h.

    Biospecimens
    Preservative Types
    • Formalin
    • Other Preservative
    • Frozen
    Diagnoses:
    • Neoplastic - Lymphoma
    Platform:
    AnalyteTechnology Platform
    DNA PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Type of fixation/preservation Formalin (buffered)
    Frozen
    Bouin's fixative
    Biospecimen Preservation Time in fixative 3 days
    4 days
    8 days
    15 days
    1 h
    2 h
    4 h
    6 h
    17 h
    24 h
    50 h
    8 h
    PCR Specific Targeted nucleic acid Framework region 1
    Framework region 2
    Framework region 3
    CK
    MBRAS
    B-globin
    Biospecimen Preservation Duration of tissue/ specimen processing Vacuum infiltration processor overnight
    Vacuum infiltration processor 48 h
    View more

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