Polymerase chain reaction denaturing gradient gel electrophoresis (PCR/DGGE)-based detection of clonal T-cell receptor γ gene rearrangements in paraffin-embedded cutaneous biopsies in cutaneous T-cell lymphoproliferative diseases

被引:10
作者
Andersen, WK [1 ]
Li, N [1 ]
Bhawan, J [1 ]
机构
[1] Boston Univ, Sch Med, Dept Dermatol, Dermatopathol Sect, Boston, MA 02118 USA
关键词
D O I
10.1111/j.1600-0560.1999.tb01825.x
中图分类号
R75 [皮肤病学与性病学];
学科分类号
100206 ;
摘要
Polymerase chain reaction (PCR)-based amplification of T-cell receptor (TCR)-gamma genes is a novel technique that can detect a clone of T cells comprising less than 1% of the total T cells in a lymphoid infiltrate(1). Besides greater sensitivity than Southern blotting, this technique can be performed with smaller quantities of lower molecular weight genomic DNA as template. We retrospectively analyzed 12 paraffin-embedded biopsies of cutaneous T-cell lymphoma (CTCL), 1 case suspicious for CTCL, 1 case of granulomatous slack skin, and 8 cases of inflammatory skin diseases to determine if PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analysis can detect TCR-gamma gene rearrangements on paraffin-embedded specimens. We were able to amplify V gamma 1-8 TCR sequences in each case and detected a dominant clone in 9 of 12 cases of CTCL and ill granulomatous slack skin. We analyzed V gamma 9 sequences in 9 cases of CTCL and detected a dominant clone in 4 cases. This study demonstrates that PCR-DGGE can easily be applied retrospectively to cutaneous biopsies of lympho-proliferative diseases when fresh tissue is not available.
引用
收藏
页码:176 / 182
页数:7
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