Application of a fluorescent PCR method for molecular diagnosis of posttransplant lymphoproliferative disorders on routine tissue sections

Elisa Gruppioni, Barbara Corti, Annalisa Altimari, Elena Gabusi, Emanuele Panza, Gian Luca Grazi, Daniele Antonio Pinna, Nicola De Ruvo, Michelangelo Fiorentino, Walter Franco Grigioni, Antonia D Errico Grigioni

Research output: Contribution to journalArticle

Abstract

Molecular detection of monoclonality can play an important role in the diagnosis of posttransplantation lymphoproliferative disorders (PTLD). To permit accurate molecular diagnosis of PTLD even on very small amounts of DNA extracted from routinely embedded histologic material, we adapted a commercially available PCR protocol (for FR-1, -2 and -3 regions), originally designed for use on fresh/frozen samples. We applied this approach on routine biopsy/surgical material of 10 PTLD (from nine patients). All three FR regions were always amplified, indicating that the extracted DNA was of medium quality. All five PTLD morphologically classified as lymphomas were monoclonal in at least one FR region. Thus, using the WHO histologic, immunohistochemical, and clinical criteria as the reference standard, the approach provided 100% sensitivity for detection of monoclonal malignancies, supporting the validity of the method. Of five specimens classified morphologically as polymorphic PTLD, three displayed a solitary IgH gene rearrangement peak, consistent with the presence of a monoclonal B-cell population (ie, monoclonal polymorphic PTLD). This rapid and straightforward procedure, which allows identification of a wide range of IgH rearrangements, could facilitate molecular analysis of PTLD in routine practice, while limiting consumption of valuable diagnostic material.

Original languageEnglish
Pages (from-to)170-176
Number of pages7
JournalDiagnostic Molecular Pathology
Volume14
Issue number3
DOIs
Publication statusPublished - Sep 2005

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Keywords

  • Immunoglobulin heavy chain genes
  • Lymphoproliferative disorders
  • Polymerase chain reaction
  • Transplantation

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

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