Nanolock-nanopore facilitated digital diagnostics of cancer driver mutation in tumor tissue

Yong Wang, Kai Tian, Ruicheng Shi, Amy Gu, Michael Pennella, Lindsey Alberts, Kent S. Gates, Guangfu Li, Hongxin Fan, Michael X. Wang, Li Qun Gu

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

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 BRAF 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 wildtype 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 BRAF 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.

Original languageEnglish (US)
Pages (from-to)975-981
Number of pages7
JournalACS Sensors
Volume2
Issue number7
DOIs
StatePublished - Jul 28 2017

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Keywords

  • BRAF
  • Diagnostics
  • Driver mutation
  • Nanolock
  • Nanopore
  • Precision medicine
  • Single molecule detection
  • Single nucleotide polymorphism (SNP)

ASJC Scopus subject areas

  • Bioengineering
  • Fluid Flow and Transfer Processes
  • Process Chemistry and Technology
  • Instrumentation

Cite this

Wang, Y., Tian, K., Shi, R., Gu, A., Pennella, M., Alberts, L., Gates, K. S., Li, G., Fan, H., Wang, M. X., & Gu, L. Q. (2017). Nanolock-nanopore facilitated digital diagnostics of cancer driver mutation in tumor tissue. ACS Sensors, 2(7), 975-981. https://doi.org/10.1021/acssensors.7b00235