Research Update CHF 02528 Final: 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
vector-borne 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, or the presence of co-infection without performing multiple assays. With the AKC-CHF support, we were able to develop a novel diagnostic tool based on high-throughput next-generation sequencing (NGS) capable of testing a large number of canine samples at once for the detection of one or more organisms in one single assay.

We were able to expand the level of specificity of the NGS when compared to traditional approaches by targeting a different gene to precisely detect each organism present in clinical specimens. We confirmed in-vitro the assay’s ability to detect low levels of infection with one organism even in the presence of high levels of other organisms.

We also evaluated a method to decrease the presence of host DNA to improve the detection of vector-borne pathogens and developed a computational pipeline to help facilitate the analysis of the sequencing data. Collectively, our efforts yielded a novel diagnostic platform for the broad detection of tick- and blood-borne organisms in dogs. The AKC-CHF support was instrumental in the development of such diagnostic tools in veterinary medicine.

Final Update – Developing a Next Generation Sequencing Diagnostic Platform for Tick-Borne Diseases