Detection of the t(14;18) translocation in frozen and formalin-fixed tissue.
As part of a retrospective study into the prevalence of the t(14;18) translocation in B-cell lymphomas, we assessed the suitability of the polymerase chain reaction (PCR) to amplify the t(14;18) major breakpoint region (MBR) in frozen and formalin-fixed tissue. Considering Southern blotting as a standard, the sensitivity of PCR was 81%. Of the various procedures used to extract DNA from paraffin-embedded tissue (PET), proteinase K digestion in the presence of nonionic detergents gave the highest yield and quality of DNA and the most efficient amplification rate. Using this method, excellent amplification rates (100%) were obtained for both the beta-globin control sequence and the MBR t(14;18) for fixed follicular lymphoma specimens collected in the previous 2 to 6 years (n = 27). Of nine older PETs, PCR on six gave inconsistent results, probably because of the poorer-quality substrate used for amplification. Specimens exposed to formol sublimate or formalin-acetic acid-alcohol were as suitable for amplification as tissues fixed in neutral-buffered formalin. The overall incidence of the MBR t(14;18) in all follicular lymphoma specimens as detected by both Southern blotting and PCR was 59% (23 of 39).
Limpens
J
J
Department of Pathology, Sylvius Laboratory, Leiden University, The Netherlands.
Beelen
M
M
Stad
R
R
Haverkort
M
M
van Krieken
J H
JH
van Ommen
G J
GJ
Kluin
P M
PM
D001483:Base Sequence; D002883:Chromosomes, Human, Pair 14; D002887:Chromosomes, Human, Pair 18; D015925:Cryopreservation; D004273:DNA, Neoplasm / Q000235:genetics; D006801:Humans; D016393:Lymphoma, B-Cell / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D016707:Tissue Fixation; D014178:Translocation, Genetic / Q000235:genetics*
D004273:DNA, Neoplasm