Thursday, October 30
7:30am Morning Coffee
8:00 Opening Comments by Session Chairperson
Nikhat Parveen, Ph.D., Associate Professor, Department of Microbiology and Molecular Genetics, Rutgers-New Jersey Medical School
8:05 Next-Generation Diagnostic Test for Lyme Disease
Benjamin Luft, M.D., Professor, Medicine, Stony Brook University
We have developed a novel sensitive and specific single-tier Lyme disease assay that has a wide range of coverage and specificity against Lyme disease including the early stages of the disease. The sensitivity of our assay far exceeds that of commercially available Lyme disease assays in patients in the early stages of the disease. Our results may lead to the development of a next-generation rapid, single-tier reference laboratory assay.
8:35 Performance of United States Serologic Assays in the Diagnosis of Lyme Borreliosis Acquired in Europe
John A. Branda, Ph.D., Microbiology Laboratory, Massachusetts General Hospital
Using serum samples from European LB patients, we compared the performance of European and US serodiagnostic tests, including newer-generation assays containing Vmp-like sequence, expressed or its C6 peptide. The sensitivity of each assay was determined using 64 serum samples from LB patients with early or late disease manifestations who acquired the infection in Europe. Specificity was measured using 100 sera from healthy subjects from a nonendemic area. European assays outperformed analogous US assays in a conventional 2-tiered testing algorithm. However, a C6 ELISA used as a stand-alone test or in the second tier of a 2-tiered algorithm performed comparably to conventional 2-tiered testing using European assays, and can be used for evaluation of any patient, regardless of travel history.
9:05 Sensitive Multiplex PCR Assay to Detect Lyme Spirochetes, Anaplasma phagocytophilum and Babesia microti
Nikhat Parveen, Ph.D., Associate Professor, Department of Microbiology and Molecular Genetics, Rutgers-New Jersey Medical School
Emergence of co-infections with tick-borne pathogens in endemic regions of the USA and Europe emphasizes the need of a single test that can detect different pathogens simultaneously. Nucleic-based diagnostic tests are becoming useful in diagnosis of active infectious diseases. We have developed a real-time multiplex PCR assay to detect the presence of three tick-borne pathogens, Borrelia burgdorferi sensulato, Babesia microti and Anaplasma phagocytophilum efficiently and accurately using the specific molecular beacon probes.
9:35 Coffee Break in the Exhibit Hall with Poster Viewing
10:05 Molecular Diagnosis of Human Anaplasmosis and Ehrlichiosis Using a Real-Time PCR Assay
Anna M. Schotthoefer, Ph.D., Marshfield Clinic Research Foundation
Human anaplasmosis and ehrlichiosis are emerging tick-borne diseases that present as non-specific, febrile illnesses, but which have the potential to be fatal. Early and accurate diagnosis and treatment are currently the most effective ways to avoid serious illness and death related to the diseases. The clinical utility of a real-time PCR assay that incorporates melt curve analysis to differentiate Anaplasma and Ehrlichia species infections will be discussed.
10:35 Immuno-PCR Detection of Lyme Disease—A Simplified and Objective Approach
Mollie W. Jewett, Ph.D., Assistant Professor, Burnett School of Biomedical Sciences, University of Central Florida College of Medicine
Immuno-PCR (iPCR) is a technique that combines ELISA-based detection specificity with the sensitivity of PCR signal amplification and has demonstrated increased sensitivity for the detection of disease biomarkers. We present evidence that iPCR is highly sensitive and specific method for the detection of Lyme disease. This is the first demonstration of iPCR for Lyme disease diagnosis and supports the replacement of two-tier testing with a more simplified and objective approach.
11:05 Sponsored Presentation (Opportunity Available)
11:35 Enjoy Lunch on Your Own
12:55 pm Comments by Session Chairperson
Chris MacManus, Ph.D., Scientist, Research and Development, Ventria Biosciences
1:00 Old Tricks and New Trends in Diagnosis of Rickettsial Diseases: From the Lab Bench to the Bedside
Marina Eremeeva, M.D., Ph.D., DSc., Associate Professor, Core Lab Director, Environmental Health Sciences, GSU
Laboratory diagnosis of classic and emerging tick-borne rickettsial diseases is an important component of patient care and surveillance. However, in most instances confirmatory diagnosis relies on less than ideal serologic assays with limited sensitivity and temporal dependence and little utility for point-of-care treatment. This presentation will focus on a concise discussion of contemporary approaches to clinically-relevant diagnoses of rickettsioses. In particular, this imperative will be articulated for both acute stage agent specific diagnostic assays and a systems biology approach to host responses and evaluating specific biomarkers of disease and exposure.
1:30 Topical and Non-Topical Treatment of Lyme Disease Shortly After a Tick Bite
Charles Pavia, Ph.D., Academic Scientist-Professor, Biomedical Sciences, NYIT College of Osteopathic Medicine and New York Medical College
We have modified the murine animal model for LD, to determine minimally effective treatment regimens, given topically or by other parenteral routes, for preventing Borrelia burgdorferi infection. Our results show that topical applications of povidone-iodine, erythromycin and tetracycline were unable to prevent borrelial dissemination to the urinary bladder or ear. On the other hand, a one or 2 dose regimen of ceftriaxone (CTX), given, i.m. or i.d. was 90-100% effective in sterilizing the urinary bladders and ear skin of the Borrelia burgdorferi-infected mice. We conclude that the topical application of various antimicrobial preparations, that are acceptable for use in humans, are unable to prevent disseminated Lyme disease, but short-course treatment with CTX, that was given by injection, is highly curative, and should be considered for further evaluation in human clinical trials.
2:00 Immunization with a Borrelia burgdorferi BB0172-Derived Peptide Protects Mice against Lyme Disease
Maria D. Esteve-Gassent, Ph.D., Assistant Professor, Department of Veterinary Pathobiology, Texas A & M University
Vaccine development efforts focused on the von Willebrand factor A domain of the borrelial protein BB0172 from which four peptides (A, B, C and D) were synthesized and conjugated to Keyhole Limpet Hemocyanin, formulated in Titer Max® adjuvant and used to immunize C3H/HeN mice subcutaneously at days 0, 14 and 21. Sera were collected to evaluate antibody responses and some mice were sacrificed for histopathology to evaluate vaccine safety. Results showed that only mice immunized with peptide B were protected against challenge with Bb. In addition, compared to the other treatment groups, peptide B-immunized mice showed very limited inflammation in the heart and joint tissues. Peptide B-specific antibody titers peaked at 8 weeks post-priming and surprisingly, the anti-peptide B antibodies did not cross-react with Bb lysates. These findings strongly suggest that peptide B is a promising candidate for the development of a new DIVA vaccine (Differentiate between Infected and Vaccinated Animals) for protection against Lyme disease.
2:30 Refreshment Break with Exhibit and Poster Viewing
3:00 Reservoir Targeted Protective Immunization against Lyme Disease (B. burgdorferi) Transmission in Mice by Oral Bait Containing ExpressTec-Made Fusion Protein Antigen
Chris MacManus, Ph.D., Scientist, Research and Development, Ventria Biosciences
Ventria Bioscience has demonstrated successful production of a recombinant Borrelia burgdorferi antigen (OspA) linked with Cholera Toxin Subunit B (CTB.rOspA) using Ventria’s ExpressTec platform for use as an oral vaccine against the spread of the Lyme disease pathogen. ExpressTec-made CTB.OspA maintains the correct conformation of the major antigenic epitope of native OspA recognized by the protective IgG monoclonal antibody, as evident in functional assays. The presence of CTB has been demonstrated to have preserved adjuvant activity as demonstrated by biochemical analysis, functional binding assays and in vivo activity. Ventria and CDC scientists have shown that ExpressTec-made CTB. OspA vaccine confers protective immunity when administered to mice as an oral bait, as measured by increased levels of protective IgG, and protection from B. burgdorferi transmission from infected ticks. ExpressTec-made CTB. OspA is being developed to disrupt the transmission cycle of the Lyme disease pathogen through baiting zoonotic reservoirs in geographic areas where Lyme is a significant threat to humans.
3:30 CASE STUDY: Blocking Pathogen Transmission at the Source: Reservoir Targeted Vaccine Against Borrelia burgdorferi
Maria Gomes-Solecki, Ph.D., Assistant Professor, Department of Microbiology, Immunology and Biochemistry, University of Tennessee Health Science Center
No human vaccine is approved by the FDA against Lyme disease and even the most efficacious human vaccines can not prevent disease expansion into other geographical areas. Alternative strategies are needed. I will provide a brief overview of the application of an OspA-based wildlife reservoir targeted vaccine aimed at reducing transmission of B. burgdorferi and present it as a strategy for reducing Lyme disease risk to humans.
4:15 Close of Conference
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