肿瘤异质性和缺乏个性化预后导致膀胱癌（BlCa）患者需要终生侵入性干预监测，凸显对现代微创工具进行疾病管理的需求。在此，我们对术前血清细胞自由DNA（cfDNA）在改善患者风险分层和预后方面的临床效用进行了评估。从190名术前BlCa患者和26名健康个体的血清样本中纯化cfDNA，并进行了两个检测：一个是自行开发的定量实时荧光PCR（qPCR）试剂，使用LEP作为参考对照，另一个是使用Qubit HS dsDNA的直接荧光法。在31个样本中进行了毛细管电泳，以进行cfDNA片段分析。非肌层侵犯性膀胱癌和肌层侵犯性膀胱癌（MIBC）的临床终点是肿瘤复发/进展和转移/死亡。毛细管电泳的cfDNA分析强调了BlCa中总和与片段相关的cfDNA水平显著增加，并与晚期疾病阶段相关。通过Qubit / qPCR评估cfDNA水平显示出高度一致的结果（rs = 0.960; P <0.001）。较高的cfDNA与MIBC和早期转移的较高风险（Qubit：危险比[HR]=3.016，P=0.009；qPCR：HR=2.918，P=0.004）和MIBC患者的不良生存（Qubit：HR=1.898，P=0.042；qPCR：HR=1.888，P=0.026）相关。多变量cfDNA拟合模型相对于疾病建立标记，为MIBC预后提供了更优异的风险分层和净益。升高的术前cfDNA水平与MIBC的短期转移风险和不良预后密切相关，支持现代无创疾病预测和管理。 ©美国临床化学协会 2023版权所有。有关授权，请发送电子邮件至： journals.permissions@oup.com。

Tumor heterogeneity and lack of personalized prognosis leads to bladder cancer (BlCa) patients' lifelong surveillance with invasive interventions, highlighting the need for modern minimally invasive tools for disease management. Herein, we have evaluated the clinical utility of preoperative serum cell-free DNA (cfDNA) in ameliorating patients' risk-stratification and prognosis.cfDNA was purified from 190 preoperative BlCa patients and 26 healthy individuals' serum samples and quantified by 2 assays: an in-house quantitative real-time PCR (qPCR) assay using LEP as reference control and a direct fluorometric assay using Qubit HS dsDNA. Capillary electrophoresis was performed in 31 samples for cfDNA fragment profiling. Tumor relapse/progression and metastasis/death were used as clinical endpoints for non-muscle-invasive bladder cancer and muscle-invasive bladder cancer (MIBC), respectively.cfDNA profiling by capillary electrophoresis highlighted that total and fragment-related cfDNA levels were significantly increased in BlCa and associated with advance disease stages. Evaluation of cfDNA levels by both Qubit/qPCR displayed highly consistent results (rs = 0.960; P < 0.001). Higher cfDNA was correlated with MIBC and stronger risk for early metastasis (Qubit:hazard ratio [HR] = 3.016, P = 0.009; qPCR:HR = 2.918, P = 0.004) and poor survival (Qubit:HR = 1.898, P = 0.042; qPCR:HR = 1.888, P = 0.026) of MIBC patients. Multivariate cfDNA-fitted models led to superior risk stratification and net benefit for MIBC prognosis compared to disease established markers.Elevated preoperative cfDNA levels are strongly associated with higher risk for short-term metastasis and poor outcome of MIBC, supporting modern noninvasive disease prognosis and management.© American Association for Clinical Chemistry 2023. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.