Tethered enzyme test for highly accurate, rapid, field-based detection of avian influenza

Highly pathogenic avian influenza (HPAI), already a major problem for poultry farms, has recently caused outbreaks in dairy cattle herds around the country, drastically reducing the viable milk output of affected cows. Furthermore, evidence shows that the virus can be transmitted to humans through contact with infected animals, raising concerns of increased spread and possible recombination with a human strain of influenza. The USDA has implemented mandatory testing for animals before crossing state lines, but testing beyond that remains extremely limited due to the turnaround time associated with the current gold standard method, RT-PCR, which requires samples to be shipped to a laboratory. A rapid, on-site test is required for farmers to make meaningful management decisions regarding the sequestering of sick animals. To address this need, TETmedical is developing a sensitive, rapid, PoC diagnostic test for HPAI H5N1 using their innovative tethered enzyme technology (TET) platform. This technology utilizes enzymes which are tethered to nanoparticles to generate a rapid viral RNA biosensor with a bioluminescent readout. The oriented immobilization of the enzymes on the nanoparticles confers advantages in speed, specificity, and sensitivity. Here, we propose to utilize sequence-specific identification oligonucleotides to target the enzymatic reaction to H5N1 RNA. A similar TET test for SARS CoV-2 RNA demonstrated high sensitivity and a 5-minute reaction time. The key advantage of this method over existing diagnostic techniques is the specificity and rapid turnaround time enabled by on-site testing, which enables immediate interventions that can reduce the spread within and between farms.