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Quantitative real-time reverse transcription-PCR assay for cyclin D1 expression: utility in the diagnosis of mantle
K E Bijwaard1, N S Aguilera, Y Monczak
1Department of Cellular Pathology, Armed Forces Institute of Pathology, 6825 16th St. NW, Washington, DC 20306-6000, USA. bijwaard@afip.osd.mil
Background:
The t(11;14)(q13;q32) translocation present in the majority of mantle cell lymphomas (MCLs) places the cyclin D1 gene under the control of immunoglobulin transcriptional regulatory elements, causing overexpression of cyclin D1. Quantification of cyclin D1 expression can distinguish MCL from other lymphomas.
Methods:
A quantitative real-time reverse transcription (RT)-PCR assay was developed for cyclin D1 mRNA suitable for use with RNA extracted from fresh and formalin-fixed, paraffin-embedded tissues. Specimens were amplified in an Applied Biosystems Model 7700 Sequence Detection System in reactions containing primers and probes for cyclin D1 and a control gene, beta(2)-microglobulin. Relative expression of the two genes was standardized against a control MCL cell line, M02058.
Results:
The range of cyclin D1 expression among 20 MCLs was substantially higher than that in other lymphomas and reactive lymph nodes. By choosing an optimal cutoff point for assessing overexpression, the sensitivity and specificity of the assay for the diagnosis of MCL in lymph node specimens both approached 100%: Overexpression was detected in 20 of 20 MCLs, but in none of 21 non-mantle-cell lymphomas or 10 reactive lymph nodes.
Conclusions:
Quantitative real-time RT-PCR for cyclin D1 overexpression provides a rapid diagnostic test with clinical utility in the diagnosis of MCL.
