title: Application of a fluorescent PCR method for molecular diagnosis of posttransplant lymphoproliferative disorders on routine tissue sections.
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.
authors.affiliation: Molecular and Transplantation Pathology Laboratory of the F. Addarii Institute of Oncology, S. Ozsola-Malpighi Hospital, University of Bologna, Bologna, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Gruppioni
authors.forename: Barbara
authors.initials: B
authors.lastname: Corti
authors.forename: Annalisa
authors.initials: A
authors.lastname: Altimari
authors.forename: Elena
authors.initials: E
authors.lastname: Gabusi
authors.forename: Emanuele
authors.initials: E
authors.lastname: Panza
authors.forename: Gian Luca
authors.initials: GL
authors.lastname: Grazi
authors.forename: Antonio Daniele
authors.initials: AD
authors.lastname: Pinna
authors.forename: Nicola
authors.initials: N
authors.lastname: De Ruvo
authors.forename: Michelangelo
authors.initials: M
authors.lastname: Fiorentino
authors.forename: Walter Franco
authors.initials: WF
authors.lastname: Grigioni
authors.forename: Antonia D'Errico
authors.initials: AD
authors.lastname: Grigioni
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D005453:Fluorescence; D015321:Gene Rearrangement; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D008223:Lymphoma / Q000235:genetics / Q000473:pathology; D008232:Lymphoproliferative Disorders / Q000175:diagnosis*; D008875:Middle Aged; D016377:Organ Transplantation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011183:Postoperative Complications / Q000175:diagnosis*; D012680:Sensitivity and Specificity
chemical_list: D007143:Immunoglobulin Heavy Chains; D004247:DNA