A vertical flow paper-microarray assay with isothermal DNA amplification for detection of Neisseria meningitidis

Talanta. 2018 Jun 1:183:192-200. doi: 10.1016/j.talanta.2018.02.070. Epub 2018 Feb 16.

Abstract

Paper-based biosensors offer a promising technology to be used at the point of care, enabled by good performance, convenience and low-cost. In this article, we describe a colorimetric vertical-flow DNA microarray (DNA-VFM) that takes advantage of the screening capability of DNA microarrays in a paper format together with isothermal amplification by means of Recombinase Polymerase Amplification (RPA). Different assay parameters such as hybridization buffer, flow rate, printing buffer and capture probe concentration were optimized. A limit of detection (LOD) of 4.4 nM was achieved as determined by tabletop scanning. The DNA-VFM was applied as a proof of concept for detection of Neisseria meningitidis, a primary cause of bacterial meningitis. The LOD was determined to be between 38 and 2.1 × 106 copies/VFMassay, depending on the choice of DNA capture probes. The presented approach provides multiplex capabilities of DNA microarrays in a paper-based format for future point-of-care applications.

Keywords: Gold nanoparticles; Microarray; Neisseria meningitidis; Paper-based biosensor; Point-of-care.

MeSH terms

  • DNA, Bacterial / analysis*
  • DNA, Bacterial / genetics
  • Neisseria meningitidis / genetics
  • Neisseria meningitidis / isolation & purification*
  • Nucleic Acid Amplification Techniques*
  • Oligonucleotide Array Sequence Analysis*
  • Paper*
  • Temperature*

Substances

  • DNA, Bacterial