Zhao Cheng, Pan Yi, Wang Yinhuai, Li Yuanwei, Han Weiqing, Lu Li, Tang Wei, Li Pei, Ou Zhenyu, Zhang Mengda, Xiong Zhuang, Xu Ran, Lu Qiang, Xu Zhenzhou, Qi Lin, Wang Long, Xu Genming
Department of Urology, the Third Xiangya Hospital, Central South University, Changsha 410013, China.
Department of Urology, Xiangya Hospital, Central South University, Changsha 410008, China.
Transl Androl Urol. 2020 Jun;9(3):1222-1231. doi: 10.21037/tau-19-774.
The clinical diagnostic method for bladder cancer is cystoscopy, an invasive, expensive and inconvenient clinical test. Using urinary cell-free DNA (cfDNA) to develop non-invasive test for bladder cancer was a promising liquid biopsy.
To improve the using rate of cfDNA template and decrease the PCR bias for liquid biopsy using urinary cfDNA, we developed a cell-free single-molecule unique primer extension resequencing (cf-SUPER) technology which was done for 29 matched urinary cfDNA and tumor DNA samples of bladder cancer patients to evaluate consistency of mutation profiles. Then, a 22 high mutational frequence genes was selected to form an uriprier panel, which was analyzed in 100 patients (47 bladder cancer cases and 53 controls) using cf-SUPER technology. This performance of the technology was evaluated using bioinformatic tools and clinical samples.
The study showed that cf-SUPER technology can accurately detect mutations with allele fractions even low as 0.01% and the DNA input as low as 1 ng. The consistency of mutation profiles and clinical pathological diagnose between 29 matched urinary cfDNA and tumor DNA samples was respectively 82.76% and 89.66% by using cf-SUPER technology. Using cf-SUPER technology, the sensitivity and specificity were 98%, 94% respectively for uriprier panel in non-invasive test.
The preliminary work shows that cf-SUPER technology will be a promising method for liquid biopsy. Focusing urinary cfDNA, the non-invasive diagnose and monitoring of bladder cancer can come true by using cf-SUPER technology.
膀胱癌的临床诊断方法是膀胱镜检查，这是一种侵入性、昂贵且不方便的临床检查。利用尿游离DNA（cfDNA）开发膀胱癌的非侵入性检测方法是一种很有前景的液体活检技术。
为了提高cfDNA模板的利用率并减少使用尿cfDNA进行液体活检时的PCR偏差，我们开发了一种游离单细胞单分子独特引物延伸重测序（cf-SUPER）技术，该技术对29例膀胱癌患者匹配的尿cfDNA和肿瘤DNA样本进行检测，以评估突变谱的一致性。然后，选择22个高突变频率基因组成一个尿液引物组，使用cf-SUPER技术对100例患者（47例膀胱癌病例和53例对照）进行分析。使用生物信息学工具和临床样本对该技术的性能进行评估。
研究表明，cf-SUPER技术能够准确检测等位基因分数低至0.01%且DNA输入量低至1 ng的突变。使用cf-SUPER技术，29例匹配的尿cfDNA和肿瘤DNA样本之间的突变谱一致性和临床病理诊断一致性分别为82.76%和89.66%。使用cf-SUPER技术，尿液引物组在非侵入性检测中的敏感性和特异性分别为98%和94%。
初步工作表明，cf-SUPER技术将是一种很有前景的液体活检方法。聚焦于尿cfDNA，利用cf-SUPER技术可以实现膀胱癌的非侵入性诊断和监测。
