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Clinical and Vaccine Immunology, September 2007, p. 1158-1164, Vol. 14, No. 9
1071-412X/07/$08.00+0     doi:10.1128/CVI.00230-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

PrimaTB STAT-PAK Assay, a Novel, Rapid Lateral-Flow Test for Tuberculosis in Nonhuman Primates{triangledown}

Konstantin P. Lyashchenko,1* Rena Greenwald,1 Javan Esfandiari,1 David Greenwald,1 Carol A. Nacy,2 Susan Gibson,3 Peter J. Didier,4 Marc Washington,5 Peter Szczerba,5 Sherri Motzel,5 Larry Handt,5 John M. Pollock,6 James McNair,6 Peter Andersen,7 Jan A. M. Langermans,8,9 Frank Verreck,9 Sean Ervin,10 Frank Ervin,10 and Candace McCombs2

Chembio Diagnostic Systems, Inc., Medford, New York,1 Sequella, Inc., Rockville, Maryland,2 University of South Alabama, Mobile, Alabama,3 Tulane National Research Primate Center, Covington, Louisiana,4 Merck Research Laboratories, West Point, Pennsylvania,5 Veterinary Sciences Division, Belfast, United Kingdom,6 Statens Serum Institut, Copenhagen, Denmark,7 Animal Science Group of Wageningen UR, Lelystad, The Netherlands,8 Biomedical Primate Research Centre, Rijswijk, The Netherlands,9 Behavioural Sciences Foundation Laboratories, Basseterre, St. Kitts,10

Received 4 June 2007/ Returned for modification 9 July 2007/ Accepted 17 July 2007

Tuberculosis (TB) is the most important zoonotic bacterial disease in nonhuman primates (NHP). The current diagnostic method, the intradermal palpebral tuberculin test, has serious shortcomings. We characterized antibody responses in NHP against Mycobacterium tuberculosis to identify immunodominant antigens and develop a rapid serodiagnostic test for TB. A total of 422 NHP were evaluated, including 243 rhesus (Macaca mulatta), 46 cynomolgus (Macaca fascicularis), and 133 African green (Cercopithecus aethiops sabaeus) monkeys at five collaborative centers. Of those, 50 monkeys of the three species were experimentally inoculated with M. tuberculosis. Antibody responses were monitored every 2 to 4 weeks for up to 8 months postinfection by MultiAntigen Print ImmunoAssay with a panel of 12 recombinant antigens. All of the infected monkeys produced antibodies at various levels and with different antigen recognition patterns. ESAT-6 and MPB83 were the most frequently recognized proteins during infection. A combination of selected antigens which detected antibodies in all of the infected monkeys was designed to develop the PrimaTB STAT-PAK assay by lateral-flow technology. Serological evaluation demonstrated high diagnostic sensitivity (90%) and specificity (99%). The highest rate of TB detection was achieved when the skin test was combined with the PrimaTB STAT-PAK kit. This novel immunoassay provides a simple, rapid, and accurate test for TB in NHP.

* Corresponding author. Mailing address: Chembio Diagnostic Systems, Inc., 3661 Horseblock Road, Medford, NY 11763. Phone: (631) 924-1135. Fax: (631) 924-6033. E-mail: klyashchenko{at}chembio.com

{triangledown} Published ahead of print on 25 July 2007.

Clinical and Vaccine Immunology, September 2007, p. 1158-1164, Vol. 14, No. 9
1071-412X/07/$08.00+0     doi:10.1128/CVI.00230-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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