Quantitation of Dihydropyrimidine Dehydrogenase Expression by Real-Time Reverse Transcription Polymerase Chain Reaction
作者： Martin R. Johnson , Kangsheng Wang , Jeffrey B. Smith , Martin J. Heslin , Robert B. Diasio
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摘要: Abstract Several recent studies have reported a correlation between intratumor dihydropyrimidine dehydrogenase (DPD) messenger RNA (mRNA) levels and sensitivity to 5-fluorouracil (5-FU). However, significant tissue requirements labor-intensive methodology limited the large-scale necessary for statistical validation. In addition, semiquantitative results obtained by these methods further limit their application. We developed real-time reverse transcription-PCR (RT-PCR) assay, based on TaqMan fluorescence methodology, capable of rapid accurate quantitation DPD mRNA in biopsy-sized samples. Results with this approach indicate linear dynamic range 108–103 copies, an intra-assay variation
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