This research will investigate technologies to enable the development of spore traps capable of in-field detection, and identification, of specific biosecurity threats.

1. biosecurity built on science
Developing tools for in-field surveillance of
pathogens
Kelly Hill
Senior Research Officer
SARDI
Plant Biosecurity Cooperative Research Centre

2. biosecurity built on science
Processing of surveillance samples - problem
Trap
Spore
Samples
Collected
from Trap
Tested in field –
kits/strips/sensors
Sent to laboratory
Protein detection
techniques
DNA extraction and
Molecular techniques
Isothermal
Amplification (LAMP,
RCA, RPA)
PCR,qPCR, dPCR
Culturing and/or
Morphological
characterisation
ELISA/ Lateral Flow
Devices
Early detection of
pathogen – alert
sent
Pathogen retained
for confirmation
analysis
Trapped spores or
molecules associated
with them
automatically
detected – real time
monitoring

3. biosecurity built on science
Investigating how to make diagnostic tests for autonomous
in-field use – components and considerations
Sample
Processing
Sample
destruction/preparation
Recognition event
Measurement of that
event
Sending information
DNA/RNA
Protein/glycoprotein/other
spore coat molecules
Specific interaction with
target e.g. DNA-DNA,
protein-protein, DNA-
protein
Sensor – Optical, Mass,
Electrochemical
Data analysis/processing
Telemetry

4. biosecurity built on science
Investigating how to make diagnostic tests for autonomous
in-field use - key output
Protein/glycoprotein/other
spore coat molecules
Specific interaction with
target e.g. DNA-DNA,
protein-protein, DNA-
protein
Sensor – Optical
Data processing
Probe Development
Platform Development
Detection of semi-intact
spore captured in solution
- Puccinia striiformis
f. sp. tritici (Pst)
(Wheat Stripe Rust)
Fiber Optic Sensor to
sensitively monitor
interactions taking place
-Fiber Optic Sensor is
spliced directly into
normal optical fibres
- Can be functionalised to
carry any probes
Dr Linh Nguyen

5. biosecurity built on science
Results so far… Probe Development
5 μg of total extracted spore coat protein per spot
(2.4mg extracted from 50mg spores, 5ug therefore
approx equiv. to 0.1mg spores)
Probe
Puccinia
striiformis
f. sp. tritici (Pst)
(Stripe Rust)
Puccinia graminis
(Stem Rust)
Dot Blot Surface Plasmon Resonance

6. biosecurity built on science
Platform Development
• The majority of label free biosensing using optical techniques is based on measuring the
refractive index change as the result of biological binding.

7. biosecurity built on science
Platform Development – An optical biosensor based on the interference
effect within an exposed core fibre
Polymer: F. M. Cox, et. al., OE,
15, 11843, 2007
Soft-glass: S. C. Warren-
Smith, et. al., OE, 17, 18533,
2009
Chalcongenide: P. Toupin, et.
al., JNS, 337, 217, 2013
Silica: R. Kostecki, et. al.,
OME, 2, 1538, 2012
Easy splicing,
durable
Fiber based label free
biosensing (e.g. all-fiber
configuration without
complex add-ons) offer
stable, alignment free,
sensitive and cost-effective
operation.

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Results so far… Platform Development
Probe

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 Refining the probe to increase sensitivity and specificity
 Testing probes on platform – Limits of Detection
 Testing accuracy and durability of sensor platform
Optimization and Validation – next steps

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 Feasibility of Approach – both probe and platform development
- Probe – different platform
- Platform – measuring refractive index changes of other indicator reactions, or
other binding partners
 Report specifying protocols to apply strategy to other targets
 A system that can be built upon with appropriate fluidics for use in an
automated capture device – spore trap
Outcome and Benefits

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 For more information, please email
Kelly.Hill3@sa.gov.au