Brucellosis is a zoonotic disease with the potential to be exploited as a bioterrorism disease. The agents of brucellosis, Brucella spp., have been classified as Category B pathogens because these bacteria are stable in the environment, easily aerosolized, highly infectious, difficult to treat, and have prolonged debilitating illness. We propose to develop and deliver novel diagnostics for brucellosis caused by Brucella melitensis. Tests to diagnose acute and chronic brucellosis are far from ideal, including issues of specificity and point- care-ruggedness and ease of use. Sensitive testing to identify pre-symptomatic patients at risk for brucellosis has never been tested. Our access to populations in Peru with a high incidence of brucellosis, both in the endemic and outbreak settings, will allow us to address these major issues of brucellosis diagnosis.
Specific Aim 1 is to develop a well-defined clinical specimen bank for validating new assays to diagnose acute brucellosis by studying human populations in a Brucella me//tens/s-endemic country. This specimen bank will be comprised of clinical specimens from acute febrile patients at high risk for brucellosis. Gold standard culture (lysis centrifugation, automated blood culture), recently developed real time PCR assays, and standard serology (Rose Bengal, slide agglutination test, tube agglutination test) will be used to diagnose brucellosis. The clinical samples (isolates, sera, buffy coats) will be aliquoted and stored under Good Clinical Practice conditions as a resource for testing the new lateral flow assays described under Specific Aim 2. The specimen bank will also be available to NIH-funded biodefense investigators for testing other modalities for brucellosis detection.
Specific aim 2 is to validate new lateral flow assays to optimize the rapidity and accuracy of brucellosis. Lateral flow assays have the advantage of being rapid, rugged, easy-to- use at point-of-care, and requiring small amounts of easily obtainable clinical sample to perform. We have already successfully used a lateral flow assay, based on Brucella LPS, to detect anti-Sruce//a antibodies in Peru and other brucellosis-endemic countries. This project proposes to transfer the technology needed to fabricate this assay to the PanBio Company (Maryland, USA and Brisbane, Australia), who will manufacture the lateral flow assay for testing on the clinical specimen bank described in Specific Aim 1 under Good Manufacturing Practice conditions. In addition, we propose to test the use of a newly characterized Brucella protein, VirB12, a protein invariably detected in small animal models of infection, in the early diagnosis of brucellosis. Finally, as an alternative to nucleic acid detection, we propose to develop a lateral flow-based antigen-detection method for the identification of Brucella infection in diverse clinical scenarios.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01AI075420-04
Application #
7932896
Study Section
Special Emphasis Panel (ZAI1-MH-M (M2))
Program Officer
Ritchie, Alec
Project Start
2007-08-01
Project End
2012-07-31
Budget Start
2010-08-01
Budget End
2011-07-31
Support Year
4
Fiscal Year
2010
Total Cost
$906,252
Indirect Cost
Name
University of California San Diego
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Cannella, Anthony P; Arlehamn, Cecilia S Lindestam; Sidney, John et al. (2014) Brucella melitensis T cell epitope recognition in humans with brucellosis in Peru. Infect Immun 82:124-31
Patra, Kailash P; Saito, Mayuko; Atluri, Vidya L et al. (2014) A protein-conjugate approach to develop a monoclonal antibody-based antigen detection test for the diagnosis of human brucellosis. PLoS Negl Trop Dis 8:e2926
Feldman, Kristyn E; Loriaux, Paul M; Saito, Mayuko et al. (2013) Ex vivo innate immune cytokine signature of enhanced risk of relapsing brucellosis. PLoS Negl Trop Dis 7:e2424
Roman, Karina; Castillo, Rosa; Gilman, Robert H et al. (2013) A foodborne outbreak of brucellosis at a police station cafeteria, Lima, Peru. Am J Trop Med Hyg 88:552-8
Agampodi, Suneth B; Moreno, Angelo C; Vinetz, Joseph M et al. (2013) Utility and limitations of direct multi-locus sequence typing on qPCR-positive blood to determine infecting Leptospira strain. Am J Trop Med Hyg 88:184-5
Cannella, Anthony P; Lin, Jennifer C; Liang, Li et al. (2012) Serial kinetics of the antibody response against the complete Brucella melitensis ORFeome in focal vertebral brucellosis. J Clin Microbiol 50:922-6
Cannella, Anthony P; Tsolis, Renee M; Liang, Li et al. (2012) Antigen-specific acquired immunity in human brucellosis: implications for diagnosis, prognosis, and vaccine development. Front Cell Infect Microbiol 2:1
Agampodi, Suneth B; Matthias, Michael A; Moreno, Angelo C et al. (2012) Utility of quantitative polymerase chain reaction in leptospirosis diagnosis: association of level of leptospiremia and clinical manifestations in Sri Lanka. Clin Infect Dis 54:1249-55
Cannella, Anthony P; Nguyen, Bichchau M; Piggott, Caroline D et al. (2011) A cluster of cutaneous leishmaniasis associated with human smuggling. Am J Trop Med Hyg 84:847-50
Liang, Li; Tan, Xiaolin; Juarez, Silvia et al. (2011) Systems biology approach predicts antibody signature associated with Brucella melitensis infection in humans. J Proteome Res 10:4813-24

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