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  Vol. 139 No. 12, December 2003 TABLE OF CONTENTS
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Improved Sensitivity of T-Cell Clonality Detection in Mycosis Fungoides by Hand Microdissection and Heteroduplex Analysis

Olivier Dereure, MD, PhD; Edi Levi, MD; Eric C. Vonderheid, MD; Marshall E. Kadin, MD

Arch Dermatol. 2003;139:1571-1575.

Background  The presence of a dominant T-cell clone is an important diagnostic criterion in cutaneous T-cell lymphomas (CTCLs) and in atypical T-cell cutaneous infiltrates. Procedures based on polymerase chain reaction (PCR) are the most sensitive to detect clonality, but heteroduplex analysis is less sensitive than other methods such as denaturing gradient gel electrophoresis.

Objective  To assess whether a gross hand microdissection of the superficial (papillary) portion of the dermal infiltrate may improve the sensitivity of T-cell clonality detection by PCR-heteroduplex analysis in CTCL.

Setting  Regional university hospital (secondary or tertiary referral center).

Patients  A total of 29 patients with a definite diagnosis of mycosis fungoides based on typical histologic and immunophenotypic features were selected with patch (16) or plaque (13) stages.

Main Outcome Measures  Assessment of T-cell clonality by PCR amplification of the T-cell receptor {gamma} chain followed by heteroduplex analysis using DNA extracted from the entire biopsy specimen and after gross microdissection of the subepidermal bandlike dermal infiltrate.

Results  T-cell clonality was demonstrated in 24 of 29 cases when hand microdissection was used compared with 16 of 29 cases with DNA analysis from entire biopsy specimens; thus, hand microdissection resulted in a sensitivity improvement of approximatively 50%.

Conclusions  Hand microdissection substantially improves the detection of a T-cell clone in CTCL when using a PCR-heteroduplex analysis and could be used routinely in the clinical evaluation of T-cell infiltrates. Importantly, the microdissection method was found to be more useful for the detection of T-cell clones in early patch stages of CTCL than in plaque-stage disease.


From the Department of Pathology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Mass (Drs Dereure, Levi, and Kadin); and the Department of Dermatology, Johns Hopkins University Medical Center, Baltimore, Md (Dr Vonderheid). The authors have no relevant financial interest in this article.



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