1 research outputs found
Nanolock–Nanopore Facilitated Digital Diagnostics of Cancer Driver Mutation in Tumor Tissue
Cancer driver mutations are clinically
significant biomarkers.
In precision medicine, accurate detection of these oncogenic changes
in patients would enable early diagnostics of cancer, individually
tailored targeted therapy, and precise monitoring of treatment response.
Here we investigated a novel nanolock–nanopore method for single-molecule
detection of a serine/threonine protein kinase gene <i>BRAF</i> V600E mutation in tumor tissues of thyroid cancer patients. The
method lies in a noncovalent, mutation sequence-specific nanolock.
We found that the nanolock formed on the mutant allele/probe duplex
can separate the duplex dehybridization procedure into two sequential
steps in the nanopore. Remarkably, this stepwise unzipping kinetics
can produce a unique nanopore electric marker, with which a single
DNA molecule of the cancer mutant allele can be unmistakably identified
in various backgrounds of the normal wild-type allele. The single-molecule
sensitivity for mutant allele enables both binary diagnostics and
quantitative analysis of mutation occurrence. In the current configuration,
the method can detect the <i>BRAF</i> V600E mutant DNA lower
than 1% in the tumor tissues. The nanolock–nanopore method
can be adapted to detect a broad spectrum of both transversion and
transition DNA mutations, with applications from diagnostics to targeted
therapy