Research Update End-yr 2 CHF 02528: Developing a Next Generation Sequencing Diagnostic Platform for Tick-Borne Diseases

Diagnostic tests based on the detection of DNA from harmful organisms in clinical samples have revolutionized veterinary medicine in the last decades. Currently, diagnostic panels for several vectorborne diseases (VBDs) are available through universities and private labs in the USA and abroad. However, the vast majority of results from sick dogs are negative, which frustrates veterinarians and dog owners trying to reach a definitive diagnosis.

These panels are based on the detection of previously known DNA sequences of each microorganism, which limits their ability to detect novel organisms. Using an innovative approach, our study proposes the adaptation of high-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. However, adaptations and optimizations of the technique are required for adequate efficiency and accuracy in detecting tick-borne bacteria. With support from the AKC Canine Health Foundation, we are investigating methods to concentrate the harmful bacteria in clinical samples, precisely detect them using NGS, and better characterize these bacteria using genetic sequences. Our most recent efforts 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. Currently, we are performing a large-scale search among billions of DNA sequences of hundreds of tick-borne organisms known to cause disease in dogs and other mammals, to find the ideal and unique “DNA fingerprint”. 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.

End-Year-2 Research Update Dr. Diniz for NGS diagnostics for TBD.