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

Abstract

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.

Microfluidics for Biomedical Analysis

“…In contrast, microfluidics devices have been adopted to measure cancer‐related biomarkers due to their remarkable features in bioanalysis. For genetic analysis of prostate cancer from patients' body fluids, an integrated “sample‐to‐targeted gene analysis” biochip was developed to simultaneously detect multiple prostate cancer‐specific genes ( TMPRSS2–ERG gene fusion, PCA3 , SChLAP1 , and KLK2 ) obtained from urine and serum liquid biopsies . The biochip consists of different functional domains of 1) lysing cells electrically for 60 s to release targeted genes, 2) conducting enhanced solid‐phase isothermal amplification via nanofluidic manipulation, 3) peroxidase‐mimicking nanozyme labeling, and 4) electrochemical target detection.…”

The Growing Impact of Micro/Nanomaterial‐Based Systems in Precision Oncology: Translating “Multiomics” Technologies

“…Reproduced with permission. [137] Copyright 2018, American Chemical Society. b) Electrochemical clamp assay with Pd nanostructured microelectrodes for the detection of ctNA.…”

The Translational Potential of Electrochemical DNA-Based Liquid Biopsy

“…Regarding the analysis of NAs present in CTCs, an analytical platform integrating electrical lysis and release of cellular targets, isothermal NA amplification and nanozyme-mediated electrochemical detection has been proposed by Trau's group (Koo et al, 2018).…”