SBIR-STTR Award

HyBeacons® Probe Assays for the Detection of Ticks and Tick-Borne Pathogens Patrick Williams, Ph.D., Evogen Inc., 8301 State Line Road, Kansas City, MO 64114 The proposed development will evaluate the use of Evogen’s HyBeacons probes for rapid, multiplexed vector and pathogen detection using Evogen ONE sample processing, multiplex amplification detection, and high temperature melt curve analysis. An Assay will be developed for Amblyomma americanum. The assays will utilizes the proprietary Evogen ONE nucleic acid extraction chemistry and Evogen’s HyBeacons probes for quantitative amplification and high resolution melt curve analysis. These unique features will make this assay superior in many applications over existing technology. The ultimate goal of this project is a deployable, integrated, automated platform of chemistries and hardware capable of rapidly detecting ticks and tick-borne pathogens. The challenge of this task is in the development and validation of HyBeacons probe assays, the validation of the Evogen ONE nucleic acid extraction chemistry, the sequential automation of the pathogen analysis via the automated sample prep (ASP), and RA-8 random access amplification modules.

Keywords:
Ticks, Hybeacons Probes, Pcr, Amblyomma Americanum, Tick-Borne Pathogens

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.

Keywords:
Tick, Vector, DNA, PCR, HyBeacons, Melt Curve, Identification