SBIR-STTR Award

In the United States there are at least eleven serious diseases that are transmitted by ticks. Current diagnostic testing methods for these disease causing pathogens are limited and/or unreliable. Furthermore, no test exists that can cover the breadth of pathogens transmitted by ticks. In our Phase I proposal, we plan to develop a tick-borne pathogen detection assay for use on the Ibis T5000 system. This system employs broad-range PCR and electrospray ionization mass spectroscopy (PCR/ESI-MS). The assay to be developed and validated in this proposal will identify and speciate broad groups of bacteria, including all groups that contain known tick- borne bacteria and flaviviruses. As our assay is broad range, any novel tick-borne bacterial/viral pathogens present will also likely be detected. To achieve our goals, we will employ three key technologies: 1) Broad- range PCR primers and ESI-MS to detect a wide variety of tick-borne pathogens that might be missed by specific PCR 2) A mechanical cell destruction method, shown to lyse even the most durable forms of bacteria, such as Bacillus anthracis spores, which may enable the identification of bacteria that previously have not been detected and increase the amount of detectable pathogen DNA 3) Whole genome amplification to increase trace levels of pathogen DNA to levels detectable by PCR. In Phase I the assay will be validated on a large number of field-collected ticks from Lyme endemic areas. In Phase II, we plan to evaluate the effectiveness of our assay on human clinical samples (e.g., erythema migrans skin biopsies, cerebral spinal fluid, synovial fluid, whole blood). Upon the completion of Phases I and II, Ibis Biosciences will make available to the healthcare community a validated assay to detect bacteria/viruses in ticks and human clinical samples. Results of this work will lead to a commercially available diagnostic to detect and identify tick-borne diseases in humans for improved medical treatment and public health surveillance. In the United States there are at least eleven serious diseases that are transmitted by ticks. Current diagnostic testing methods for these disease causing pathogens are limited and/or unreliable. Furthermore, no test exists that can cover the breadth of pathogens transmitted by ticks. In this Phase I SBIR proposal we will develop and validate a broad-range tick-borne pathogen detection assay using field collected ticks. In Phase II we will continue this work using human clinical samples. Results of this work will lead to a commercially available diagnostic to detect and identify tick-borne diseases in humans for improved medical treatment and public health surveillance.

Public Health Relevance:
This Public Health Relevance is not available.

Thesaurus Terms:
There Are No Thesaurus Terms On File For This Project.

There is an increasing number of cases and spread of Lyme and other tick-borne diseases in the United States. The same tick that can transmit the agent of Lyme disease, Borrelia burgdorferi, can simultaneously transmit other pathogens such as Babesia, Anaplasma, and Rickettsia. This is of great concern to the public and their physicians. This concern is reflected in a greater demand for tests that can provide information as to what is in an individual tick and what can be transmitted. Current diagnostic testing methods for these disease causing pathogens are generally limited to a single microbe and/or unreliable. No test exists that can cover the breadth of pathogens transmitted by ticks. In our Phase I application, we achieved our aim, the proof of concept, that our methodology could detect multiple microbes and even in a single tick. That system employs broad-range PCR and mass spectroscopy (PCR/ESI-MS). The assay identified and speciated broad groups of bacteria, including known tick- borne bacteria and flaviviruses. The success of this, coupled with the increasing demand and increasing number of searches performed for Borrelia alone, is the basis for Phase 2 to concentrate on the development of a tick-borne pathogen detection assay using the high throughput PCR/ESI-MS. In Phase 1, the proof of principle of detection of pathogens was accomplished with 16 primer pairs in 16 wells of a 96 well kitted plate. In Phase 2, Goal 1 of this is to increase clinically useful throughput time and efficiency of the assay and consequently reduce costs. We will have a target goal of 8 wells (PCR reactions) by eliminating redundancy and multiplexing. Goal 2: Optimize and add to these, Babesia detection primers. Goal 3: Create a replenishable bank of nucleic acid extraction from infected ticks for future investigation of this type of assay as well as tick-human interface research. This bank will be of commercial value. Upon the completion of Phase2, we will be in a position to provide a tick-microbial diagnostic service for researchers, physicians, public health entities, private and individual citizen sectors. This will be an important tool in management of Lyme and related tick-borne diseases for improved medical treatment and public health surveillance. It will also be the foundation for doing the same in patient skin, blood, joint fluid, and cerebrospinal fluid.

Public Health Relevance:
The ability to accurately identify multiple pathogens in the ticks capable of causing Lyme disease and related disorders will be important in management strategy of patients exposed or bitten by ticks. It will help reduce rising costs to the patient and the public health system.

Thesaurus Terms:
Alphaproteobacteria; Anaplasma; Anthrax Spore; Arbovirus, Group B; Articulation; Assay; B. Anthracis Spore; B. Burgdorferi; B.Burgdorferi; Babesia; Bacillus Anthracis Spore; Bacteria; Bartonella; Bioassay; Biologic Assays; Biological Assay; Bite; Blood; Blood Volume; Borrelia; Borrelia Burgdorferi; Borrelia Burgdorferi Sensu Stricto; Businesses; Cells; Cerebrospinal Fluid; Clinical; Coupled; Cowdria; Coxiella; Cytolysis; Dna; Deoxyribonucleic Acid; Detection; Development; Diagnostic; Diagnostic Services; Diagnostic Tests; Disease; Disorder; Ehrlichia; Family; Flavivirus; Foundations; Francisella; Future; Gammaproteobacteria; Gene Products, Rna; Genome; Genomics; Goals; Health System; Human; Human, General; Incidence; Individual; Investigation; Investigators; Ixodes; Ixodes (Genus); Ixodes Tick; Ixodida; Joints; Liquid Substance; Lyme Borreliosis; Lyme Disease; Lyme Disease Spirochete; Lysis; Man (Taxonomy); Man, Modern; Mass Spectrum; Mass Spectrum Analysis; Mechanics; Medical; Method Loinc Axis 6; Methodology; Methods; Microbe; Modeling; Mollicutes; Nucleic Acids; Order Spirochaetales; Organism; Pathogen Detection; Pathway Interactions; Patients; Peptides; Phase; Photometry/Spectrum Analysis, Mass; Physicians; Piroplasma; Population Surveillance; Position; Positioning Attribute; Protocol; Protocols Documentation; Protozoa; Protozoal; Public Health; Public Health Surveillance; Rna; Rna, Non-Polyadenylated; Reaction; Reference Standards; Research; Research Personnel; Research Specimen; Researchers; Resistance; Reticuloendothelial System, Blood; Ribonucleic Acid; Rickettsia; Rochalimaea; Sampling; Skin; Specimen; Spectrometry, Mass; Spectroscopy, Mass; Spectrum Analyses, Mass; Spectrum Analysis, Mass; Spirochaetales; Spirochetes; Spiroplasma; System; System, Loinc Axis 4; Testing; Tick-Borne Diseases; Ticks; Time; United States; Validation; Alpha Proteobacteria; Anthracis Spore; Base; Borrelial; Clinical Relevance; Clinically Relevant; Cost; Diagnosis Service; Disease/Disorder; Fluid; Gamma Proteobacteria; Improved; Instrument; Liquid; Living System; Lyme Spirochete; Meetings; Member; Microbial; Pathogen; Pathway; Public Health Medicine (Field); Public Health Relevance; Resistant; Spinal Fluid; Success; Tool; Vector