Point-of-care assays are easy-to-use, portable and inexpensive tests that can
be used to aid diagnostics by measuring levels of disease-specific molecules
in settings where access to advanced laboratory equipment and trained
personnel are limited, such as at the patient's bedside or in low resource
parts of developing countries. In order to achieve high multiplexing
capacities, such assays can be based on planar microarrays consisting of
spots immobilized on a flat surface or on particle-based microarrays based
on populations of encoded particles. The aim of the work presented in this
thesis is to develop new point-of-care amenable planar and particle-based
microarrays that allow for highly multiplexed assays while maintaining low
sample-to-result times, complexity and instrumentation requirements.
Paper I demonstrates the use graphically encoded particles for colorimetric
detection of autoantibodies using a consumer-grade flatbed scanner. Four
graphical characters on the surface of each particle allows for millions of
codes and the use of gold nanoparticles as a detection label allows both the
code and the signal intensity to be read out in a single image.
Paper II describes a signal enhancement method that increases the
sensitivity of gold nanoparticle detection on planar microarrays. Using this
method, detection of allergen-specific IgE can be carried out using a
consumer-grade flatbed scanner instead of a more expensive fluorescence
scanner without sacrificing assay performance.
Paper III demonstrates the use of an isothermal DNA amplification method
for detection of adenoviral DNA on a paper-based microarray. Using an
isothermal amplification method eliminates the need for a thermocycler,
reducing the instrumentation required for such detection.
Paper IV shows the use of solid-phase PCR to amplify bacterial DNA directly
on the surface of particles. This strategy reduces assay time by eliminating
the need for separate amplification and hybridisation steps.