Background:Molecular diagnostics has become a critical component of precision oncology in solid tumors, including colorectal cancer, yet the use of formalin-fixed, paraffin-embedded (FFPE) tissue often suffers from DNA degradation that compromises sequencing quality. This study aimed to evaluate the feasibility and effectiveness of using fresh, intraoperatively collected tumor tissue for next-generation sequencing-based molecular diagnostics in colorectal cancer.Methods:Tissue samples from 24 patients undergoing colorectal tumor resection were obtained based on macroscopic evaluation and tested with a custom gene panel. Sequencing metrics, mutation profiles, and correlations with clinical and pathological features were analyzed.Results:All samples yielded high-quality sequencing data. Oncogenic or likely oncogenic variants were detected in 21 out of 24 samples (87.5%), predominantly affecting genes frequently involved in colorectal cancer carcinogenesis, includingAPC,TP53, andKRAS. In three cases, no typical mutations were found despite visual confirmation of tumor tissue during surgery, which may be attributed to insufficient tumor cellularity or molecular alterations beyond the panel’s scope.Conclusions:The results support the use of fresh tissue as a high-quality source for molecular diagnostics, capable of reducing turnaround time and avoiding formalin-induced artifacts. However, the findings also highlight the diagnostic risk of relying solely on macroscopic tumor assessment without histological confirmation. Overall, fresh tissue-based testing represents a promising yet currently investigational approach that can enhance molecular diagnostics in colorectal cancer.
背景：分子诊断已成为实体瘤（包括结直肠癌）精准肿瘤学的关键组成部分，但使用福尔马林固定、石蜡包埋（FFPE）组织常因DNA降解而影响测序质量。本研究旨在评估使用术中新鲜采集的肿瘤组织进行基于新一代测序的结直肠癌分子诊断的可行性与有效性。 方法：基于肉眼评估获取24例接受结直肠肿瘤切除术患者的组织样本，并使用定制基因组合进行检测。分析测序指标、突变谱及其与临床病理特征的相关性。 结果：所有样本均获得高质量测序数据。24例样本中有21例（87.5%）检出致癌或可能致癌的变异，主要涉及结直肠癌发生中常见的基因，包括APC、TP53和KRAS。有3例样本虽在术中肉眼确认肿瘤组织，但未发现典型突变，这可能归因于肿瘤细胞含量不足或基因组合覆盖范围外的分子改变。 结论：研究结果支持将新鲜组织作为分子诊断的高质量来源，能够缩短检测周期并避免福尔马林诱导的人工假象。然而，结果也凸显了仅依赖肉眼肿瘤评估而未经组织学确认的诊断风险。总体而言，基于新鲜组织的检测是一种具有前景但目前仍处于研究阶段的方法，可提升结直肠癌的分子诊断水平。
Evaluating the Utility of Fresh Tissue in Molecular Diagnostics of Colorectal Cancer
肿瘤(癌症)患者之家