CVI Accepts, published online ahead of print on 4 March 2009

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Clin. Vaccine Immunol. doi:10.1128/CVI.00038-09
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Highly accurate antibody assays for early and rapid detection of tuberculosis in African and Asian elephants

Rena Greenwald, Olena Lyashchenko, Javan Esfandiari, Michele Miller, Susan Mikota, John H. Olsen, Ray Ball, Genevieve Dumonceaux, Dennis Schmitt, Torsten Moller, Janet B. Payeur, Beth Harris, Denise Sofranko, W. Ray Waters, and Konstantin P. Lyashchenko^*

Chembio Diagnostic Systems, Inc., Medford, NY, USA; Disney's Animal Programs, Lake Buena Vista, FL, USA; Elephant Care International, USA; Busch Gardens Tampa Bay, Tampa, FL, USA; Missouri State University, Springfield, MO, USA; Kolmarden Zoo and Wildlife Park, Kolmarden, Sweden; United States Department of Agriculture, Animal and Plant Health Inspection Service, National Veterinary Services Laboratories, Ames, IA, USA; United States Department of Agriculture, Animal and Plant Health Inspection Service, Animal Care, Fort Collins, CO, USA; United States Department of Agriculture, National Animal Disease Center, Ames, IA, USA

^* To whom correspondence should be addressed. Email: klyashchenko{at}chembio.com.

Tuberculosis (TB) in elephants is a re-emerging zoonotic disease caused primarily by Mycobacterium tuberculosis. Current methods for screening and diagnosis rely on trunk wash culture, which has serious limitations due to low test sensitivity, slow turn-around time, and variable sample quality. Innovative and more efficient diagnostic tools are urgently needed. We describe three novel serologic techniques, ElephantTB STAT-PAK kit, multiantigen print immunoassay (MAPIA), and dual path platform (DPP) VetTB test, for rapid antibody detection in elephants. The study was performed with serum samples from 236 captive African and Asian elephants from 53 different locations in the US and Europe. The elephants were divided into three groups based on disease status and history of exposure: 1) 26 animals with culture-confirmed TB due to M. tuberculosis or M. bovis, 2) 63 exposed elephants from known infected herds which had never produced a culture positive result from trunk wash samples, and 3) 147 elephants without clinical symptoms suggestive of TB, with consistently negative trunk wash culture results, and with no history of potential exposure to TB in the past 5 years. Elephants with culture-confirmed TB and a proportion of exposed but trunk wash culture negative elephants produced robust antibody responses to multiple antigens of M. tuberculosis, with seroconversions detectable years before TB positive cultures obtained from trunk wash specimens. ESAT-6 and CFP10 proteins were immunodominant antigens recognized by elephant antibodies during disease. The serologic assays demonstrated 100% sensitivity and 95-100% specificity. Rapid and accurate antibody tests to identify infected elephants will likely allow earlier and more efficient treatment, thus limiting transmission of infection to other susceptible animals and humans.