Application of a fluorescent PCR method for molecular diagnosis of posttransplant lymphoproliferative disorders on routine tissue sections.
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.
Gruppioni
Elisa
E
Molecular and Transplantation Pathology Laboratory of the F. Addarii Institute of Oncology, S. Ozsola-Malpighi Hospital, University of Bologna, Bologna, Italy.
Corti
Barbara
B
Altimari
Annalisa
A
Gabusi
Elena
E
Panza
Emanuele
E
Grazi
Gian Luca
GL
Pinna
Antonio Daniele
AD
De Ruvo
Nicola
N
Fiorentino
Michelangelo
M
Grigioni
Walter Franco
WF
Grigioni
Antonia D'Errico
AD
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
D007143:Immunoglobulin Heavy Chains; D004247:DNA