Premium
Improved detection of clonality in cutaneous T‐cell lymphomas using laser capture microdissection
Author(s) -
Yazdi Amir S.,
Medeiros L. Jeffrey,
Puchta Ursula,
Thaller Eva,
Flaig Michael J.,
Sander Christian A.
Publication year - 2003
Publication title -
journal of cutaneous pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.597
H-Index - 75
eISSN - 1600-0560
pISSN - 0303-6987
DOI - 10.1034/j.1600-0560.2003.00099.x
Subject(s) - laser capture microdissection , mycosis fungoides , biopsy , polymerase chain reaction , microdissection , dna extraction , pathology , lymphoma , population , skin biopsy , cutaneous t cell lymphoma , biology , medicine , gene , gene expression , biochemistry , environmental health
Background: The diagnosis of cutaneous T‐cell lymphoma is a challenge for both the pathologist and the clinician. This is particularly true for distinguishing early‐stage mycosis fungoides from dermatitis. In this clinical setting, the presence of a clonal T‐cell population supports lymphoma. Methods: Usually, routinely processed paraffin‐embedded material is available for gene rearrangement analysis, and polymerase chain reaction (PCR)‐based methods to assess clonality can be performed. One drawback of this approach is that sensitivity is suboptimal in biopsy specimens in which the lymphocytic infiltrate represents only a small percentage of all cells present. Another drawback is that DNA extraction from routinely processed, paraffin‐embedded tissue is a time‐consuming and labor‐intensive procedure which can take up to 5 days in our laboratory. To bypass these problems, we used laser capture microdissection (LCM) to obtain lymphocytic infiltrates from tissue sections of formalin‐fixed, paraffin‐embedded skin biopsy specimens. This approach allows for more specific PCR assessment of the lymphocytic infiltrate and for rapid DNA extraction and PCR analysis. Results: Using the LCM approach, we could demonstrate clonal T‐cell receptor γ gene rearrangements in biopsy specimens that did not show clonality using DNA extracted by conventional methods from full tissue sections. In addition, DNA extraction and PCR analysis can be performed in 11 h. Conclusion: In conclusion, applying LCM to clonality analysis of cutaneous lymphocytic infiltrates is rapid and more sensitive than conventional methods, and we recommend introducing this approach into the routine diagnostic setting.