Detection of Khapra Beetle Environmental DNA Using Portable Technologies in Australian Biosecurity

Khapra beetle, Trogoderma granarium Everts, 1898, is a serious pest of stored grain products globally. Environmental DNA (eDNA)-based methods offer sensitive detection tools used to inform biosecurity officers on the presence high-risk pests. This study tested laboratory and portable molecular technologies detect khapra beetle environmental extracted from dust samples collected during responses (Tuggeranong Fyshwick) incursions in Australia. Airborne floor were opportunistically using handheld vacuum cleaners eDNA was either field or laboratory-based extraction analyzed benchtop real time PCR machines with two TaqMan one LAMP-based assay. We successfully collected, extracted, amplified by qPCR, but failed amplify T. LAMP. The Laboratory qPCR machine showed significantly higher mean Ct values ( p < 0.001) positive detections for both assays compared thermocycler. yield kits vacuumed airborne (Mean ± S.D. = 5.52 4.45 4.77 1.68 ng/μL, respectively), kits, (1.75 1.17 1.36± 1.29 ng/μL samples, respectively). There no significant differences between collection amplification associated platform this study. Portable (Franklin™ Real Time Thermocycler Genie III) accurately all tissue derived assay optimisation testing, highlighting capacity these complement confirming specimen ID. high incidence negative controls 12.3 % Fyshwick 50 %), mostly attributed use contaminated cleaners. discuss suitable minimize sample cross-contamination, potential as applications, suitability eDNA-based global trade most invasive important pests worldwide.