在肿瘤学中，药物基因组学的应用已成为例行临床实践的一部分。尤其是对于接受氟嘧啶和伊立替康治疗的患者，脱氢脱氢嘧啶酶（DPYD）和UDP葡萄糖醛酸转移酶（UGT1A1）的基因型检测对于治疗管理至关重要。我们实验室在肿瘤学中药物基因诊断服务方面的独特方法是将两种实时聚合酶链式反应方法相结合：LightSNiP检测法（TIB MOLBIOL）和更近期的荧光共振能量转移（FRET）探针技术（核医学激光）以及TaqMan检测法（Thermo Fisher）用于通过第二次提取的DNA确认变异等位基因的存在。我们发现，FRET和LightSNiP检测法在熔解曲线分析中的检测优势超过了竞争性TaqMan技术。当使用特定突变的TaqMan检测法时，可能会错过意外的基因变异，但所获得的信息可用于在治疗后出现意外毒性时进行调整。TaqMan和FRET检测法的结合帮助我们实现了更准确的基因分型和对患者的正确判断。DPYD FRET检测法的附加价值在于，在所述指南中详细列出的多态性的扩增剖面中，还可以检测到c.2194G>A (*6 rs1801160)，这在发生严重毒性时被引用为需要调查的变异。至于UGT1A1（TA）n启动子多态性（rs3064744），FRET检测法的独特且积极的特点是可以清晰地识别所有潜在的变异等位基因，包括在非洲裔美国人中频繁出现的（TA）5和（TA）8等位基因。我们的临床实践强调了不仅需要快速且易于使用的检测法，如新的FRET检测法，而且需要准确和全面的基因分型来进行良好的药物基因诊断活动。© 2023 Montrasio et al.

The application of pharmacogenetics in oncology is part of the routine clinical practice. In particular, genotyping of dihydropyrimidine dehydrogenase (DPYD) and UDP-glucuronosyltransferase (UGT1A1) is crucial to manage the treatment of patients taking fluoropyrimidines and irinotecan. The unique approach of our laboratory to the pharmacogenetic diagnostic service in oncology is to combine two real-time PCR methods, LightSNiP assay (TIB MOLBIOL), and more recently FRET (Fluorescent Resonance Energy Transfer) probes technology (Nuclear Laser Medicine), plus TaqMan assay (Thermo Fisher) for the confirmation of the presence of variant alleles on DNA from a second extraction. We found that both the FRET and LightSNiP assays, where detection occurs by melting curve analysis, offer an advantage over the competing TaqMan technology. Whereas unexpected genetic variants may be missed using a mutation-specific TaqMan assay, the information thus obtained can be useful to adjust the therapy in case of unexpected post-treatment toxicity. The combination of TaqMan and FRET assays helped us to achieve more accurate genotyping and a correct result for the patient. The added value of the DPYD FRET assay is the possibility of detecting, with the same amplification profile of the polymorphisms detailed in the guidelines, also the c.2194G>A (*6 rs1801160), cited in the recommendations as a variant to be investigated in case of severe toxicity. Regarding the UGT1A1 (TA)n promoter polymorphism (rs3064744), the distinctive and positive feature of the FRET assay is to allow clearly identifying all those potential variant alleles, including the (TA)5 and (TA)8 alleles, that are frequent in African Americans. Our clinical practice emphasizes the importance of not only rapid and easy-to-use assays, such as the new FRET ones, but also of accurate and comprehensive genotyping for good pharmacogenetic diagnostic activity.© 2023 Montrasio et al.