DETECTION OF THE T(14 18) TRANSLOCATION IN FROZEN AND FORMALIN-FIXED TISSUE

被引:30
作者
LIMPENS, J
BEELEN, M
STAD, R
HAVERKORT, M
VANKRIEKEN, JHJM
VANOMMEN, GJB
KLUIN, PM
机构
[1] LEIDEN UNIV, SYLVIUS LAB, DEPT PATHOL, 2300 RA LEIDEN, NETHERLANDS
[2] LEIDEN UNIV, SYLVIUS LAB, DEPT HUMAN GENET, 2300 RA LEIDEN, NETHERLANDS
关键词
POLYMERASE CHAIN REACTION; PARAFFIN-EMBEDDED TISSUE; FIXATION; DNA EXTRACTION; FOLLICULAR LYMPHOMA; BCL-2; T(14 18);
D O I
10.1097/00019606-199303000-00015
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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).
引用
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页码:99 / 107
页数:9
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