Tuberculosis (TB) is the most important zoonotic bacterial disease in nonhuman primates (NHP).Tuberculosis (TB) remains one of the most important bacterial diseases of nonhuman primates (NHP) because of its frequency of occurrence, ability to spread rapidly, high mortality rates, and zoonotic potential (14,26,32). The disease in NHP is caused by the same organisms that are responsible for TB in humans (Mycobacterium tuberculosis) and cattle (Mycobacterium bovis) (4,11,33,35). Infection can be readily transmitted from one monkey to another or from monkeys to humans and vice versa (26,31).To control the spread of this devastating illness, captive NHP are screened for TB regularly. The USDA-approved method tests for delayed hypersensitivity in response to an intradermal injection of mammalian old tuberculin (MOT) into the skin of the eyelid (intrapalpebral tuberculin test) (8). MOT is composed of poorly defined mycobacterial antigens that are known to be highly cross-reactive (12, 25). The visual scoring of the test is largely subjective, often allowing confusion between negative and positive results (6, 15). In spite of rigorous testing and import quarantine measures, outbreaks of TB in NHP continue to occur in research laboratories, resulting in the destruction of numerous valuable animals in the United States and elsewhere (14, 32). Many healthy primates are also euthanized unnecessarily because of false-positive skin test reactions that might be due to a nonspecific inflammatory response to MOT or because of sensitization with cross-reactive nonpathogenic species of environmental mycobacteria (10,15,17,19). Thus, alternative diagnostic approaches for antemortem TB testing of NHP are urgently needed.We have recently developed two membrane-based antibody detection methods, MultiAntigen Print ImmunoAssay (MAPIA) and a lateral-flow test, and demonstrated the potential for rapid TB serodiagnosis in various host species (16,23,34). In the present study, we characterized the antibody responses in NHP experimentally infected with M. tuberculosis and developed the PrimaTB STAT-PAK assay, a novel lateral-flow test for antibody detection in TB. This test was evaluated in comparison with the intradermal palpebral tuberculin test and other laboratory diagnostic methods over the course of experimental infection.
MATERIALS AND METHODS
Animals and infection.A total of 422 NHP were used, including 243 rhesus (Macaca mulatta), 46 cynomolgus (Macaca fascicularis), and 133 African green (Cercopithecus aethiops sabaeus) monkeys from five collaborative centers (Table 1). At four research sites, experimental infection with M. tuberculosis was performed on 50 animals, including 29 rhesus, 16 cynomolgus, and 5 African green monkeys. Animal infection experiments were performed in biosafety level 3 facilities according to the study protocols approved by the Institutional Animal Care and Use Committees. After prestudy quarantine and extensive evaluation for TB, monkeys were inoculated intratracheally with various doses and strains of M. ...