Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR/DGGE)-based detection of clonal T-cell receptor gamma gene rearrangements in paraffin-embedded cutaneous biopsies in cutaneous T-cell lymphoproliferative diseases.
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. 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 Vgamma1-8 TCR sequences in each case and detected a dominant clone in 9 of 12 cases of CTCL and in granulomatous slack skin. We analyzed Vgamma9 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 lymphoproliferative diseases when fresh tissue is not available.
Andersen
W K
WK
Department of Dermatology, Boston University School of Medicine, Massachusetts, USA.
Li
N
N
Bhawan
J
J
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015703:Antigens, CD / Q000032:analysis; D001706:Biopsy; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015334:Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor; D019675:Genes, T-Cell Receptor gamma / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D016410:Lymphoma, T-Cell, Cutaneous / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008875:Middle Aged; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012867:Skin / Q000737:chemistry / Q000378:metabolism / Q000473:pathology; D012878:Skin Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology
D015703:Antigens, CD; D017931:DNA Primers; D004273:DNA, Neoplasm