Development of a Molecular Assay to Identify Ticks that Vector Rickettsial Diseases

Tick borne diseases pose a significant threat to deployed military forces and local civilian populations. Different species of ticks vector different pathogens; therefore it is important to be able to differentiate numerous species of ticks at any stage of their life cycle. Evogen has optimized the extraction and purification of tick DNA from whole tick bodies and individual tick legs. This purified DNA has been tested against primer and probe designs to 16S rDNA and the ITS2 gene targets. Based on these results, additional gene targets have been selected for evaluation: COI, COII, COB, 12 S RNA, and the mt Control Region. Using representative ticks from North America, these gene targets will be amplified, sequenced, and analyzed to facilitate the design of molecular assays to perform species level identification. HyBeacons probes will be designed for each of the gene targets providing the ability to perform qPCR and high resolution melt curve analysis. A deployable tick assay will provide clinicians with the ability to conduct rapid and accurate surveillance of the vector population of ticks and provide an additional estimate of human encounters with infected ticks. This information will support significant efforts to protect military forces and civilian populations