Department of Physiology & Biochemistry, Faculty of Medicine and Surgery, University of Malta, Malta.
Department of Pathology, Faculty of Medicine & Surgery, University of Malta, Malta.
Methods. 2019 Apr 1;158:86-91. doi: 10.1016/j.ymeth.2018.10.008. Epub 2018 Oct 22.
Patient stratification, prognosis and disease monitoring are three important aspects of personalized cancer medicine. With traditional serum tumour protein biomarkers showing lack of specificity and sensitivity, and tumour heterogeneity affecting the response to targeted therapy based on tissue biomarkers, the focus has shifted to the use of molecular tumour signatures as specific biomarkers. Multiplex microsphere-based panels are robust and cost-effective, high throughput molecular assays, which can accurately characterize tumours even from small amounts of poor quality nucleic acids. Only few studies have reported the use of microspheres (beads) to quantify RNA expression of targets of interest simultaneously (multiplexing). This review is an overview of the various applications of bead-based RNA panels in molecular oncology, with focus on the Invitrogen™ QuantiGene™ Plex Assay (Thermo Fisher Scientific), and provides a comparison with PCR-based and other methodologies. The advantages of multiplex bead assays are exemplified by the quantification of RNA expression in formalin-fixed, paraffin embedded (FFPE) archival tissue and the simultaneous detection of biomarkers in low input samples, including quantification of markers in microdissected tissue material, to characterise heterogeneous tumour sites within a sample, and by the detection of markers in low numbers of circulating tumour cells.
患者分层、预后和疾病监测是个性化癌症医学的三个重要方面。由于传统的血清肿瘤蛋白生物标志物特异性和敏感性不足,且肿瘤异质性影响基于组织生物标志物的靶向治疗反应,因此研究重点已转向使用分子肿瘤特征作为特异性生物标志物。基于多重微球的面板是强大且具有成本效益的高通量分子检测方法,即使是从小量质量较差的核酸中,也能准确地对肿瘤进行特征分析。只有少数研究报告了使用微球(珠)同时定量感兴趣的靶标 RNA 表达(多重检测)。本综述概述了基于微球的 RNA 面板在分子肿瘤学中的各种应用,重点介绍了 Invitrogen™ QuantiGene™ Plex 检测法(Thermo Fisher Scientific),并与基于 PCR 的方法和其他方法进行了比较。多重微球检测的优势在于可对福尔马林固定石蜡包埋(FFPE)存档组织中的 RNA 表达进行定量,以及可对低输入样本中的生物标志物进行同时检测,包括对微切割组织材料中的标志物进行定量,以对样本中的异质性肿瘤部位进行特征分析,以及对低数量循环肿瘤细胞中的标志物进行检测。