Kretschmer-Kazemi Far Rosel, Frank Kirsten, Sczakiel Georg
Institut für Molekulare Medizin, Universität zu Lübeck and UKSH, Campus Lübeck, Ratzeburger Allee 160, 23538 Lübeck, Germany.
Cancers (Basel). 2021 Aug 30;13(17):4381. doi: 10.3390/cancers13174381.
Body fluids in the context of cancer diagnosis are the primary source of liquid biopsy, i.e., biomarker detection that includes blood and serum, urine, and saliva. RNA represents a particular class of biomarkers because it is thought to monitor the current status of gene expression in humans, in organs, and if present, also in tumors. In case of bladder cancer, we developed a scheme that describes, in detail, all steps from the collection of urine samples from patients, stabilization of samples, their transportation, storage, and marker analysis by qPCR-based technology. We find that urine samples prepared according to this protocol show stability of RNA over more than 10 days at unchilled temperatures during shipping. A specific procedure of primer design and amplicon evaluation allows a specific assignment of PCR products to human genomics and transcriptomics data collections. In summary, we describe a technical option for the robust acquisition of urine samples and the quantitative detection of RNA-based tumor markers in case of bladder cancer patients. This protocol is for general use, and we describe that it works for any RNA-based tumor marker in urine of cancer patients.
在癌症诊断中，体液是液体活检的主要来源，即生物标志物检测，包括血液、血清、尿液和唾液。RNA代表一类特殊的生物标志物，因为它被认为可以监测人类、器官以及（如果存在的话）肿瘤中的基因表达现状。对于膀胱癌，我们制定了一个方案，详细描述了从患者尿液样本采集、样本稳定、运输、储存到通过基于qPCR的技术进行标志物分析的所有步骤。我们发现，按照该方案制备的尿液样本在运输过程中未冷藏的温度下，RNA能在10多天内保持稳定。引物设计和扩增子评估的特定程序允许将PCR产物明确对应到人类基因组学和转录组学数据集中。总之，我们描述了一种在膀胱癌患者中稳健获取尿液样本并定量检测基于RNA的肿瘤标志物的技术方法。该方案可普遍使用，并且我们表明它适用于癌症患者尿液中任何基于RNA的肿瘤标志物。
