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Temperature gradient gel electrophoresis for analysis of a polymerase chain reaction‐based diagnostic clonality assay in the early stages of cutaneous T‐cell lymphomas
Author(s) -
Menke Martin A. O. H.,
Tiemann Markus,
Vogelsang Dörthe,
Boie Christina,
Parwaresch Reza
Publication year - 1995
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.11501601119
Subject(s) - mycosis fungoides , agarose gel electrophoresis , polymerase chain reaction , microbiology and biotechnology , temperature gradient gel electrophoresis , gel electrophoresis , biology , multiplex polymerase chain reaction , agarose , multiplex , lymphoma , dna , gene , genetics , immunology , 16s ribosomal rna
By means of a multiplex polymerase chain reaction (PCR) we amplified rearranged T‐cell receptor γ chain genes to detect monoclonality in 370 formalin‐fixed skin biopsy specimens, showing histological features of parapsoriasis or mycosis fungoides. PCr products were analyzed by temperature gradient gel electrophoresis (TGGE). We selected 20 positive cases for use in a comparison of this technique with conventional agarose and polyacrylamide gel electrophoresis (PAGE). With TGGE the T‐cells had shown monoclonality in 272 of the 370 cases; with agarose electrophoresis they did so in only 5 of the 20 selected cases and with PAGE in 16. Where multiple biopsy specimens from the same patient were analyzed, PCR products showed identical rearrangement patterns in TGGE. TGGE is an efficient technique that works on routine material and can help to verify a histological diagnosis of cutaneous T‐cell lymphoma.