Research Update CHF 02528 MY3: Developing a Next Generation Sequencing Diagnostic Platform for Tick-Borne Diseases

Despite the wide availability of tick-borne disease panels at laboratories in the USA and abroad, most results from sick dogs are negative, which frustrates veterinarians and dog owners trying to reach a definitive diagnosis.

Using an innovative approach, our study proposes the adaptation of high[1]throughput next-generation sequencing (NGS) to the detection of tick-borne bacteria in dog blood to overcome the limitations of the current diagnostics. Our previous results showed the potential of NGS in detecting known and potentially novel tick-borne bacteria. Recently, we have tested a commercially available kit designed to concentrate microbial DNA from blood to facilitate the detection of bacteria in circulation. However, even when 24 times more blood volume was used, no evidence of the concentration of blood microbiome or tick-borne organisms was detected using two distinct molecular assays.

As microbial concentration remains the next logical step in improving the detection of tick-borne organisms, we continue to investigate other potential approaches. In parallel, we have focused on finding new and better “DNA fingerprints” of these harmful bacteria, so we can precisely detect them from blood samples of naturally-infected dogs.

Unfortunately, the COVID-19 pandemic forced us to stop all benchtop activities related to this effort. With new biosecurity measures implemented in our campus, we plan to resume our efforts, and tests in the lab the DNA fingerprint candidates that we previously identified using large-scale bioinformatic analyses. Collectively, our efforts may yield a novel diagnostic platform for the broad detection of tick- and blood-borne organisms in dogs. The AKC-CHF support has been instrumental in the development of such diagnostic tools in veterinary medicine.

Mid-Year-3 Research Update Dr. Diniz Next Generation testing for TBD.