端粒酶逆转录酶启动子 (TERTp) 突变经常是靶向肿瘤标志物，然而，它们存在于 GC 含量高的区域，这在使用简单的分子技术甚至下一代测序 (NGS) 进行检查时提出了挑战。在膀胱癌 (BC) 中，TERTp 突变特别频繁，然而，没有任何可用工具能够证明通过简单的非侵入性技术检测 TERTp 突变的有效性。因此，我们开发了一种基于 PCR 的新型方法来检测两种最常见的 TERTp 突变，并证明了其在 BC 样本分析中的用途。开发的 SHARD-PCR TERTp 突变检测技术需要 PCR 和限制性消化步骤，即使在设备较差的实验室中也可以轻松实施。具有已知突变状态的细胞系用于方法开发。对 BC 患者的匹配尿液和肿瘤组织样本进行了分析，并通过下一代测序验证了结果。通过 SHARD-PCR 对 18 个尿液和相应的肿瘤组织样本进行分析，结果显示样本对完美匹配，这与相应的 NGS 结果相似：14 个样本在 -124 位点显示突变，2 个样本在 -146 位点显示突变，无突变在两个样品中均被检测到。我们的研究作为概念验证，受到样本量较小的限制，但它表明 SHARD-PCR 是一种简单、经济且高度可靠的检测 TERTp 突变的方法，TERTp 突变在不同癌症类型中常见。对于膀胱癌，SHARD-PCR 可以使用非侵入性样本进行，并且可以替代或补充当前使用的技术。© 2024。作者。

Telomerase reverse transcriptase promoter (TERTp) mutations are frequently targeted tumor markers, however, they reside in regions with high GC content, which poses challenges when examined with simple molecular techniques or even with next-generation sequencing (NGS). In bladder cancer (BC), TERTp mutations are particularly frequent, however, none of the available tools have demonstrated efficacy in detecting TERTp mutations via a simple noninvasive technique. Therefore, we developed a novel PCR-based method for the detection of the two most common TERTp mutations and demonstrated its use for the analysis of BC samples. The developed SHARD-PCR TERTp mutation detection technique requires PCR and restriction digestion steps that are easily implementable even in less well-equipped laboratories. Cell lines with known mutational status were utilized for method development. Matching urine and tumor tissue samples from BC patients were analyzed, and the results were validated by next-generation sequencing. Analysis of eighteen urine and corresponding tumor tissue samples by SHARD-PCR revealed perfect matches in sample pairs, which paralleled the corresponding NGS results: fourteen samples exhibited mutations at the -124 position, two samples showed mutations at the -146 position, and no mutations were detected in two samples. Our study serves as a proof-of-concept and is limited by its small sample size, nonetheless, it demonstrates that SHARD-PCR is a simple, economic and highly reliable method for detecting TERTp mutations, which are common in different cancer types. For bladder cancer, SHARD-PCR can be performed with the use of noninvasive samples and could replace or complement currently used techniques.© 2024. The Author(s).