A Sample-to-Targeted Gene Analysis Biochip for Nanofluidic Manipulation of Solid-Phase Circulating Tumor Nucleic Acid Amplification in Liquid Biopsies

The
use of circulating tumor nucleic acids (ctNA) in patient liquid
biopsies for targeted genetic analysis is rapidly increasing in clinical
oncology. Still, the call for an integrated methodology, which is
both rapid and sensitive for analyzing trace ctNA amount in liquid
biopsies, has unfortunately not been fully realized. Herein, we performed
complex liquid biopsy sample-to-targeted genetic analysis on a biochip
with a 50 copies-detection limit within 30 min. Our biochip uniquely
integrated the following: (1) electrical lysis and release of cellular
targets with minimal processing; (2) nanofluidic manipulation to accelerate
molecular kinetics of solid-phase isothermal amplification; and (3)
single-step capture and amplification of multiple NA targets prior
to nanozyme-mediated electrochemical detection. Using prostate cancer
liquid biopsies, we successfully demonstrated multifunctionality for
cancer risk prediction; correlation of serum and urine analyses; and
cancer relapse monitoring.

循环肿瘤核酸（circulating tumor nucleic acids, ctNA）在患者液体活检中用于靶向基因分析的应用，在临床肿瘤学中正快速增长。然而，针对可快速且高灵敏地分析液体活检中痕量ctNA的集成化分析方法的需求，目前尚未得到充分满足。本研究中，我们依托一款检测限达50拷贝、可在30分钟内完成检测的生物芯片，完成了从复杂液体活检样本到靶向基因分析的全流程检测。该生物芯片创新性集成了三项核心技术：(1) 仅需极少量处理步骤的电裂解与细胞靶标释放；(2) 纳米流体操控以加速固相等温扩增的分子动力学过程；(3) 在纳米酶介导的电化学检测前，完成多核酸靶标的单步捕获与扩增。利用前列腺癌液体活检样本，我们成功验证了该平台的多功能性，其可应用于癌症风险预测、血清与尿液分析相关性研究及癌症复发监测等场景。