Surface-Modified Mesh Filter for Direct Nucleic Acid Extraction and its Application to Gene Expression Analysis

Adv Healthc Mater. 2017 Oct;6(20). doi: 10.1002/adhm.201700642. Epub 2017 Jul 17.

Abstract

Rapid and convenient isolation of nucleic acids (NAs) from cell lysate plays a key role for onsite gene expression analysis. Here, this study achieves one-step and efficient capture of NA directly from cell lysate by developing a cationic surface-modified mesh filter (SMF). By depositing cationic polymer via vapor-phase deposition process, strong charge interaction is introduced on the surface of the SMF to capture the negatively charged NAs. The NA capturing capability of SMF is confirmed by X-ray photoelectron spectroscopy, fluorescent microscopy, and zeta potential measurement. In addition, the genomic DNAs of Escherichia Coli O157:H7 can be extracted by the SMF from artificially infected food, and fluorescent signal is observed on the surface of SMF after amplification of target gene. The proposed SMF is able to provide a more simplified, convenient, and fast extraction method and can be applied to the fields of food safety testing, clinical diagnosis, or environmental pollutant monitoring.

Keywords: cationic polymers; initiated chemical vapor deposition (iCVD); nucleic acid extraction; pathogen detection.

MeSH terms

  • DNA, Bacterial / analysis
  • DNA, Bacterial / isolation & purification*
  • Environmental Monitoring
  • Escherichia coli O157 / genetics*
  • Genome, Bacterial
  • Limit of Detection
  • Microscopy, Electron, Scanning
  • Photoelectron Spectroscopy
  • Polymers / chemistry*
  • Solid Phase Extraction / instrumentation
  • Solid Phase Extraction / methods*
  • Spectrum Analysis, Raman
  • Surface Properties

Substances

  • DNA, Bacterial
  • Polymers