We present a workflow using fluorescent color-coded Luminex beads to detect the
outcome of droplet PCR assay. The assay was performed to detect three important
poultry pathogens: avian influenza, infectious laryngotracheitis virus and
campylobacter. Droplet-based TaqMan PCR has been commonly used for detection
of rare and significant biomarkers in clinical samples. However, the spectral overlap
of fluorescent TaqMan probes limits the detection to 5 different targets in a single
assay. The color codes of the Luminex detection beads allowed accurate classification
of the different bead sets used in this assay concurrently. The target-specific capture
probes coupled to distinct bead sets enabled capture and detection of target DNA in
the droplet. The capture assay detected target DNA of all three poultry pathogens with
high specificity, from samples with average target concentration of 1 template per
droplet. This workflow demonstrates that the detection panel of droplet PCR assay
can be increased to potentially detect multiple targets in a sample by utilizing the
scalability offered by the color-coded detection beads.