Clonality testing of lymphoproliferative disorders in a large cohort of primary and consultant biopsies.
BACKGROUND
Lymphoproliferative disease often presents the clinician and pathologist with a diagnostic dilemma, particularly in the early course of the disease.


METHODS
We used modified BIOMED-2 protocols to detect monoclonal expansions of immunoglobulin heavy chain (IgH) and T-cell receptor (TCR) genes in 957 formalin-fixed paraffin-embedded samples from 717 patients. To eliminate false-positive results, heteroduplex analysis was used after PCR reactions. The impact of different fixatives on DNA quality and performance of PCR was assessed.


RESULTS
In the class of B lymphomas we detected clonal IgH rearrangement in nearly 80% of cases and in the class of T lymphomas in 64% of cases. Performance of the assays was 94.7% and 92.5% for IgH and TCR clonality, respectively. Clonality rates in various B and T lymphomas were in concordance with previous studies. We also present 10 difficult cases where PCR analysis of IgH and TCR gene rearrangements significantly contributed to a decision on the correct diagnosis.


CONCLUSIONS
These results confirm that the PCR-based analysis is suitable as a routine method and is helpful in establishing a diagnosis in morphologically unclear cases.
Department of Clinical and Molecular Pathology, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc, Czech Republic.
Michaela
M
Svachova
Department of Clinical and Molecular Pathology, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc, Czech Republic.
Martin
M
Tichy
Department of Clinical and Molecular Pathology, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc, Czech Republic.
Patrik
P
Flodr
Department of Clinical and Molecular Pathology, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc, Czech Republic.|Institute of Molecular and Translation Medicine, Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Republic.
Jana
J
Steigerova
Department of Clinical and Molecular Pathology, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc, Czech Republic.|Institute of Molecular and Translation Medicine, Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Republic.
Zdenek
Z
Kolar
Department of Clinical and Molecular Pathology, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc, Czech Republic.|Institute of Molecular and Translation Medicine, Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Republic.
Jan
J
Bouchal
D000328:Adult; D000368:Aged; D001706:Biopsy; D002999:Clone Cells; D015331:Cohort Studies; D005260:Female; D015321:Gene Rearrangement; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics*; D008232:Lymphoproliferative Disorders / Q000000981:diagnostic imaging* / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D055815:Young Adult
D007074:Immunoglobulin G; D011948:Receptors, Antigen, T-Cell
clonality; IgH and TCR gene rearrangement; lymphoid proliferation; polymerase chain reaction; BIOMED-2