[Study of B-lymphocyte clonality using in vitro gene amplification (PCR) in paraffin embedded samples].
Family-specific primers were used in polymerase chain reaction (PCR) to analyse clonality of immunoglobulin heavy chain (IgH) gene rearrangement. DNA templates were extracted from formalin-fixed paraffin-embedded biopsies from B-cell lymphomas (n = 19), T-cell lymphomas (n = 3), reactive lymphadenopathies (n = 20) and negative controls (n = 2). PCR was also performed on DNA extracted from fragments of the same biopsies that were either fixed in Bouin's solution (n = 24) or snap-frozen (n = 22). The latter were also studied for IgH gene rearrangement by the Southern blot technique. Polyclonal or clonal fragments were obtained from all frozen biopsies. No amplification was observed in 3 out of 44 formalin fixed specimens and 18 out of 24 Bouin fixed specimens. A clonal amplification was only observed in 16 out of 19 formalin-fixed B-cell lymphoma specimens. Therefore this technique allows to study B-cell clonality from formalin-fixed, embedded material.
Dubus
P
P
Service d'Anatomie Pathologique, CHU de Bordeaux, Pessac.
Labouyrie
E
E
Bilhou-Nabera
C
C
Belleannee
G
G
Vergier
B
B
Bernard
P
P
de Mascarel
A
A
Merlio
J P
JP
D000085:Acetates; D019342:Acetic Acid; D001483:Base Sequence; D001706:Biopsy; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D005404:Fixatives; D005557:Formaldehyde; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain; D006801:Humans; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D016612:Paraffin Embedding; D010851:Picrates; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results
D000085:Acetates; D005404:Fixatives; D010851:Picrates; D005557:Formaldehyde; C026239:Bouin's solution; D004247:DNA; D019342:Acetic Acid
Etude de la clonalité B lymphocytaire par la technique d'amplification de gène in vitro (PCR) à partir de prélèvements fixés et inclus en paraffine.