Double signal amplification strategy for ultrasensitive electrochemical biosensor based on nuclease and quantum dot-DNA nanocomposites in the detection of breast cancer 1 gene mutation

摘要

Rapid and efficient detection of microRNA (miRNA) of breast cancer 1 gene mutation (BRCA1) at their earliest stages is one of the crucial challenges in cancer diagnostics. In this study, a highly-sensitive electrochemical DNA biosensor was fabricated by double signal amplification (DSA) strategy for the detection of ultra-trace miRNA of BRCA1. In the presence of target miRNA of BRCA1, the well-matched RNA-DNA duplexes were specifically recognized by double-strand specific nuclease (DSN), and the DNA part of the duplexes were then cleaved and miRNAs were released to trigger another following cycle, which produced a primarily amplified signal by such a cyclic enzymatic signal amplification (CESA). Then triple-CdTe quantum dot labelled DNA nanocomposites (3QD@DNA NC) was selectively hybridized with the cleaved DNA probe on the electrode and produced multiply amplified signals. The biosensor exhibited a high sensitivity for the detection of miRNA of BRCA1 in concentrations ranging from 5 aM to 5 fM, and its detection limit of 1.2 aM was obtained, which is two or three orders of magnitude lower than those by single signal amplification strategy such as CESA or QD-labeled DNA probes. The as-prepared biosensor was successfully used to detect the miRNA of BRCA1 in human serum samples with acceptable stability, good reproducibility, and good recovery. The proposed DNA biosensor based on double signal amplification strategy provided a feasible, rapid, and sensitive platform for early clinical diagnosis and practical applications.